CA3073185A1

CA3073185A1 - Treatment of inflammatory disease using ingestible device to release immune modulator

Info

Publication number: CA3073185A1
Application number: CA3073185A
Authority: CA
Inventors: Mitchell Lawrence Jones; Christopher Loren WAHL; Sharat Singh
Original assignee: Progenity Inc
Current assignee: Biora Therapeutics Inc
Priority date: 2017-08-15
Filing date: 2018-08-13
Publication date: 2019-02-21
Also published as: US20200323772A1; WO2019036382A1; CN111225686A; EP3668544A1

Abstract

This disclosure features methods and compositions for treating inflammatory disorders or conditions that arise in a tissue originating from the endoderm using an immune modulator.

Description

DEMANDE OU BREVET VOLUMINEUX
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PLUS D'UN TOME.

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2 TREATMENT OF INFLAMMATORY DISEASE USING INGESTIBLE
DEVICE TO RELEASE IMMUNE MODULATOR
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims priority to U.S. provisional patent application serial numbers 62/545,894 filed on August 15, 2017, 62/583,969 filed on November 9, 2017, 62/596,041 filed on December 7, 2017, 62/599,000 filed December 14, 2017, 62/599,005 filed December 14, 2017, and 62/650,900 filed on March 30, 2018, the contents of each of which is hereby incorporated by reference in its entirety into this application.
TECHNICAL FIELD
This disclosure features methods and compositions for treating a disease or condition in a tissue originating from the endoderm.
BACKGROUND
The tissues that originate from the endoderm are linked by, e.g., a lymphatic system.
For example, the gastrointestinal tract, gallbladder, pancreas, and liver (all of which originate from the endoderm) drain into the mesenteric lymph system. Although the tissues that originate from the endoderm are succeptible to different inflammatory diseases or conditions, immune modulators that preferentially suppress immune response of the mesenteric lymph system may represent a new way to treat inflammatory diseases or conditions of tissues that arise from the endoderm.
SUMMARY
The present invention is based on the discovery that local and/or topical delivery of an immune modulator to the gastrointestinal tract significantly reduced the mean number of pro-inflammatory T cells found locally within the mesenteric lymph nodes when compared to systemic and vehicle treatment. In addition, there were fewer a4137-expressing T cells found in adjacent inflamed tissues proximal (small intestinal Payer's Patches) to where the drug was delivered (cecum).
The traditional immune modulator mechanism of action for systemically administered immune modulators is a systemic blockage of immune cell activation (e.g., T-cell activation), a systemic decrease in the secretion and/or expression of pro-inflammatory cytokines, and/or a systemic increase in the secretion of anti-inflammatory cytokines (e.g., systemically blocking T cell surface a4137 integrin/MAdCAM-1 interaction, which leads to thereby reduced trafficking to inflamed tissues). However, when an immune modulator was applied topically (e.g., locally) to the gastrointestinal system (using any of the devices described herein), a significant, profound, and unexpected reduction in T cell number was observed in inflamed tissues, draining lymph nodes, as well as tissues adjacent and upstream of the topical site of drug delivery. These results suggest that blocking local a4137 integrin interactions and T cell recruitment may be responsible. It is possible that blocking local a4137 integrin interactions and T cell recruitment using immune modulators, may be reducing immune cell trafficking or reducing the "imprinting" of T cells to express a4137 and become "gut homing." It is possible that topically-applied immune modulators are moving in the extracellular or lymph spaces including from distal to proximal gut. It is also possible that reduced trafficking of these immune cells through the lymph structures is resulting in reduced levels of immune cells in tissues that are not in areas directly treated with an immune modulator.
The observation of the pharmacodynamics effects of gastrointestinal-delivered immune modulators extend to the mesenteric lymph nodes (MSN), and the organs and tissues that drain into the MSN (a tissue originating from the endoderm), which suggests that locally-delivered (gastrointestinal tissue-delivered) immune modulators may have anti-inflammatory effects for a range of indications beyond the site of delivery. In some embodiments, the compositions and methods of the present invention may be used to treat diseases and conditions that arise in a tissue originating from the endoderm. The endoderm forms the gastrointestinal tract, respiratory tract, endocrine glands and organs, auditory system and urinary system; therefore, the present invention includes compositions and methods for treating diseases and conditions found in the following tissues: the stomach, the colon, the liver, the pancreas, the gallbladder, the urinary bladder, the epithelial parts of trachea, the lungs, the pharynx, the thyroid, the parathyroid, the intestines, and the gallbladder.
Provided herein are methods of treating an inflammatory disease or condition that arrises in a tissue originating from the endoderm in a subject, that include:
releasing an immune modulator at a location in the gastrointestinal tract of the subject, where the methods include administering to the subject a pharmaceutical composition includes a therapeutically effective amount of the immune modulator.

In some embodiments of these methods, the pharmaceutical composition is an ingestible device and the method includes administering orally to the subject the pharmaceutical composition. In some embodiments of these methods, the method does not include releasing more than 10% of the immune modulator at a location that is not proximate to the intended site of release. In some embodiments of these methods, the method provides a concentration of the immune modulator at a location that is an intended site of release that is 2-100 times greater than at a location that is not the intended site of release.
In some embodiments of any of the methods described herein, the method provides a concentration of the immune modulator in the plasma of the subject that is less than 3 [tg/mL, less than 0.3 [tg/mL, or less than 0.01 [tg/mL.
In some embodiments of any of the methods described herein, the metho provides a C24 value of the immune modulator in the plasma of the subject that is less than 3 [tg/mL, less than 0.3 [tg/mL, or less than 0.01 [tg/mL.
In some embodiments of any of the methods described herein, the immune modulator is an inhibitory nucleic acid. In some embodiments of any of the methods described herein, the immune modulator is a small molecule. In some embodiments of any of the methods described herein, the immune modulator is an antisense nucleic acid. In some embodiments of any of the methods described herein, the immune modulator is a ribozyme. In some embodiments of any of the methods described herein, the immune modulator is a siRNA.
In some embodiments of any of the methods described herein, the immune modulator is present in a pharmaceutical formulation within the device. In some embodiments of any of the methods described herein, the formulation is a solution of the immune modulator in a liquid medium. In some embodiments of any of the methods described herein, the formulation is a suspension of the immune modulator in a liquid medium.
In some embodiments of any of the methods described herein, the tissue originating from the endoderm is selected from the group of: the stomach, the colon, the liver, the pancreas, the urinary bladder, the epithelial parts of the trachea, the lungs, the pharynx, the thyroid, the parathyroid, the intestines, and the gallbladder. In some embodiments of any of the methods described herein, the inflammatory disease or condition originating from the .. endoderm is selected from the group of: gastritis, Celiac disease, hepatitis, alcoholic lever disease, fatty liver disease (hepatic steatosis), non-alcoholic fatty liver disease (NASH), cirrhosis, primary schlerosing cholangitis, pancreatitis, insterstitial cystitits, asthma, chronic obstructic pulmonary disease, pulmonary fibrosis, pharyngitis, thyroiditis, hyperthyroidism,

3 parathyroiditis, nephritis, Hashimoto's disease, Addison's disease, Graves' disease, Sjogren syndrome, type 1 diabetes, pelvic inflammatory disease, auditory canal inflammation, tinnitus, vestibular neuritis, otitis media, auditory canal inflammation, tracheitis, cholestatic liver disease, primary biliary schlerosis, liver parenchyma, an inherited metabolic disorder of the liver, Byler syndrome, cerebrotendinous, xanthomatosis, Zellweger's syndrome, neonatal hepatitis, cystic fibrosis, ALGS (Alagilles syndrome), PFIC (progressive familial intrahepatic cholestasis), autoimmune hepatitis, primary biliary cirrhosis (PBC), liver fibrosis, NAFLD, portal hypertension, general cholestasis, such as in jaundice due to drugs or during pregnancy, intra- and extrahepatic cholestasis, such as hereditary forms of cholestasis, such as PFIC1, gall stones and choledocholithiasis, malignancy causing obstruction of the biliary tree, symptoms (scratching, pruritus) due to cholestasis/jaundice, chronic autoimmune liver disease leading to progressive cholestasis, and pruritus of cholestatic liver disease, duodenal ulcers, enteritis (radition-, chemotherapy-, or infection-induced enteritis), diverticulitis, pouchitis, cholecystitis, and cholangitis. In some embodiments of any of the methods described herein, the inflammatory disease or condition that arises in a tissue originating from the endoderm is inflammation of the liver.
In some embodiments of any of the methods described herein, the immune modulator is released at a location in the large intestine of the subject. In some embodiments of any of the methods described herein, the location is in the proximal portion of the large intestine. In some embodiments of any of the methods described herein, the location is in the distal portion of the large intestine.
In some embodiments of any of the methods described herein, the immune modulator is released at a location in the ascending colon of the subject. In some embodiments of any of the methods described herein, the location is in the proximal portion of the ascending colon. In some embodiments of any of the methods described herein, the location is in the distal portion of the ascending colon.
In some embodiments of any of the methods described herein, the immune modulator is released at a location in the cecum of the subject. In some embodiments of any of the methods described herein, the location is in the proximal portion of the cecum. In some embodiments of any of the methods described herein, the location is in the distal portion of the cecum.
In some embodiments of any of the methods described herein, the immune modulator is released at a location in the sigmoid colon of the subject. In some embodiments of any of

4 the methods described herein, the location is in the proximal portion of the sigmoid colon. In some embodiments of any of the methods described herein, the location is in the distal portion of the sigmoid colon. In some embodiments of any of the methods described herein, the immune modulator is released at a location in the transverse colon of the subject. In some embodiments of any of the methods described herein, the location is in the proximal portion of the transverse colon. In some embodiments of any of the methods described herein, the location is in the distal portion of the transverse colon.
In some embodiments of any of the methods described herein, the immune modulator is released at a location in the descending colon of the subject. In some embodiments of any of the methods described herein, the location is in the proximal portion of the descending colon. In some embodiments of any of the methods described herein, the location is in the distal portion of the descending colon.
In some embodiments of any of the methods described herein, the immune modulator is released at a location in the small intestine of the subject. In some embodiments of any of .. the methods described herein, the location is in the proximal portion of the small intestine. In some embodiments of any of the methods described herein, the location is in the distal portion of the small intestine.
In some embodiments of any of the methods described herein, the immune modulator is released at a location in the duodenum of the subject. In some embodiments of any of the methods described herein, the location is in the proximal portion of the duodenum. In some embodiments of any of the methods described herein, the location is in the distal portion of the duodenum.
In some embodiments of any of the methods described herein, the immune modulator is released at a location in the jejunum of the subject. In some embodiments of any of the .. methods described herein, the location is in the proximal portion of the jejunum. In some embodiments of any of the methods described herein, the location is in the distal portion of the jejunum.
In some embodiments of any of the methods described herein, the immune modulator is released at a location in the ileum of the subject. In some embodiments of any of the .. methods described herein, the location is in the proximal portion of the ileum. In some embodiments of any of the methods described herein, the location is in the distal portion of the ileum.

5 In some embodiments of any of the methods described herein, the location at which the immune modulator is released is 10 cm or less from an intended site of release. In some embodiments of any of the methods described herein, the location at which the immune modulator is released is 5 cm or less from an intended site of release. In some embodiments of any of the methods described herein, the location at which the immune modulator is released is 2 cm or less from an intended site of release.
In some embodiments of any of the methods described herein, the immune modulator is released by mucosal contact. In some embodiments of any of the methods described herein, the immune modulator is delivered to the location by a process that does not comprise systemic transport of the immune modulator.
Some embodiments of any of the methods described herein further include identifying an intended site of release of the immune modulator using a method that includes imaging of the gastrointestinal tract. In some embodiments of any of the methods described herein, the method includes identifying an intended site of release of the immune modulator, prior to .. administering the pharmaceutical composition. In some embodiments of any of the methods described herein, the method includes releasing the immune modulator substantially at the same time as identifying the intended site of release of the immune modulator.
In some embodiments of any of the methods described herein, the methods include (a) identifying a subject having an inflammatory disease or condition that arises in a tissue originating from the endoderm, and (b) evaluating the subject for suitability to treatment.
In some embodiments of any of the methods described herein, the releasing of the immune modulator is triggered by one or more of: a pH in the jejunum from 6.1 to 7.2, a pH
in the mid small bowel from 7.0 to 7.8, a pH in the ileum from 7.0 to 8.0, a pH in the right colon from 5.7 to 7.0, a pH in the mid colon from 5.7 to 7.4, or a pH in the left colon from 6.3 to 7.7, such as 7Ø
In some embodiments of any of the methods described herein, the releasing of the immune modulator is not dependent on the pH at or in the vicinity of the location.
In some embodiments of any of the methods described herein, the releasing of the immune modulator is triggered by degradation of a release component located in the device.
In some embodiments of any of the methods described herein, the releasing of the immune modulator is not triggered by degradation of a release component located in the device. In some embodiments of any of the methods described herein, the releasing of the immune modulator is not dependent on enzymatic activity at or in the vicinity of the location. In some

6 embodiments of any of the methods described herein, the releasing of the immune modulator is not dependent on bacterial activity at or in the vicinity of the location.
In some embodiments of any of the methods described herein, the composition includes a plurality of electrodes including a coating, and releasing the immune modulator is triggered by an electric signal by the electrodes resulting from the interaction of the coating with an intended site of release of the immune modulator. In some embodiments of any of the methods described herein, the release of the immune modulator is triggered by a remote electromagnetic signal.
In some embodiments of any of the methods described herein, the release of the immune modulator is triggered by generation in the composition of a gas in an amount sufficient to expel the immune modulator. In some embodiments of any of the methods described herein, the release of the immune modulator is triggered by an electromagnetic signal generated within the device according to a pre-determined drug release profile.
In some embodiments of any of the methods described herein, the ingestible device includes an ingestible housing, wherein a reservoir storing the immune modulator is attached to the housing. Some embodiments of any of the methods described herein further include:
detecting when the ingestible housing is proximate to an intended site of release, where releasing the immune modulator includes releasing the therapeutically effective amount of the immune modulator from the reservoir proximate the intended site of release in response to the detection. In some embodiments of any of the methods described herein, the detecting includes detecting via one or more sensors coupled to the ingestible housing.
In some embodiments of any of the methods described herein, the one or more sensors include a plurality of coated electrodes and wherein detecting includes receiving an electric signal by one or more of the coated electrodes responsive to the one or more electrode contacting the respective intended site of release. In some embodiments of any of the methods described herein, the releasing includes opening one or more valves in fluid communication with the reservoir. In some embodiments of any of the methods described herein, the one or more valves is communicably coupled to a processor positioned in the housing, the processor communicably coupled to one or more sensors configured to detect the intended site of release. In some embodiments of any of the methods described herein, the releasing includes pumping the therapeutically effective amount of the immune modulator from the reservoir via pump positioned in the ingestible housing. In some embodiments of the methods described herein, the pump is communicably coupled to a processor positioned in the housing, the processor communicably coupled to one or more sensors configured to detect an intended site

7 of release of the immune modulator. In some embodiments of any of the methods described herein, the therapeutically effective amount of the immune modulator is stored in the reservoir at a reservoir pressure higher than a pressure in the gastrointestinal tract of the subject.
Some embodiments of any of the methods described herein further include anchoring the ingestible housing at a location proximate to the intended site of release in response to the detection. In some embodiments of any of the methods described herein, the anchoring the ingestible housing includes one or more legs to extend from the ingestible housing.
In some embodiments of any of the methods described herein, the amount of the immune modulator that is administered is from about 1 mg to about 500 mg. In some embodiments of any of the methods described herein, the immune modulator is an antibody or an antigen-binding antibody fragment. In some embodiments of any of the methods described herein, the antibody is a humanized antibody.
In some embodiments, the subject is administered the dose of the immune modulator once a day. In some embodiments, the subject is administered the dose of the immune modulator once every two days.
In some embodiments of any of the methods described herein, the amount of the immune modulator is less than an amount that is effective when the immune modulator is administered systemically. In some embodiments of any of the methods described herein, the methods include administering (i) an amount of the immune modulator that is an induction dose. Some embodiments of any of the methods described herein further include (ii) administering an amount of the immune modulator that is a maintenance dose following the administration of the induction dose. In some embodiments of any of the methods described herein, the induction dose is administered once a day. In some embodiments of any of the methods described herein, the induction dose is administered once every two days. In some embodiments of any of the methods described herein, the induction dose is administered once every three days. In some embodiments of any of the methods described herein, the induction dose is administered once a week. In some embodiments of any of the methods described herein, step (ii) is repeated one or more times. In some embodiments of any of the methods described herein, step (ii) is repeated once a day over a period of about 6-8 weeks.
In some embodiments of any of the methods described herein, step (ii) is repeated once every three days over a period of about 6-8 weeks. In some embodiments of any of the methods described herein, step (ii) is repeated once a week over a period of about 6-8 weeks.

8 In some embodiments of any of the methods described herein, the induction dose is equal to the maintenance dose. In some embodiments of any of the methods described herein, the induction dose is greater than the maintenance dose. In some embodiments of any of the methods described herein, the induction dose is 5 times greater than the maintenance dose. In some embodiments of any of the methods described herein, the induction dose is 2 times greater than the maintenance dose.
In some embodiments of any of the methods described herein, the method includes releasing the immune modulator at the location in the gastrointestinal tract as a single bolus.
In some embodiments of any of the methods described herein, the method includes releasing the immune modulator at the location in the gastrointestinal tract as more than one bolus. In some embodiments of any of the methods described herein, the method includes delivering the immune modulator at the location in the gastrointestinal tract in a continuous manner. In some embodiments of any of the methods described herein, the method includes delivering the immune modulator at the location in the gastrointestinal tract over a time period of 20 or more minutes. In some embodiments of any of the methods described herein, the method does not include delivering an immune modulator rectally to the subject. In some embodiments of any of the methods described herein, the method does not include delivering an immune modulator via an enema to the subject. In some embodiments of any of the methods described herein, the method does not include delivering an immune modulator via suppository to the subject. In some embodiments of any of the methods described herein, the method does not include delivering an immune modulator via instillation to the rectum of the subject. In some embodiments of any of the methods described herein, the method does not include surgical implantation.
In some embodiments of any of the methods described herein, the immune modulator is an IL-12/IL-23 inhibitor. In some embodiments of any of the methods described herein, the immune modulator is a TNFa inhibitor. In some embodiments of any of the methods described herein, the immune modulator is a IL-6 receptor inhibitor. In some embodiments of any of the methods described herein, the immune modulator is a CD40/CD4OL
inhibitor.
In some embodiments of any of the methods described herein, the immune modulator is a IL-1 inhibitor. In some embodiments of any of the methods described herein, the immune modulator is a PDE4 inhibitor.
In some embodiments of any of the methods described herein, the composition is an autonomous device. In some embodiments of any of the methods described herein, the

9 composition includes a mechanism capable of releasing the immune modulator. In some embodiments of any of the methods described herein, the composition includes a tissue anchoring mechanism for anchoring the composition to the location. In some embodiments of any of the methods described herein, the tissue anchoring mechanism is capable of activation for anchoring to the location. In some embodiments of any of the methods described herein, the tissue anchoring mechanism includes an osmotically-driven sucker. In some embodiments of any of the methods described herein, the tissue anchoring mechanism includes a connector operable to anchor the composition to the location. In some embodiments of any of the methods described herein, the connector is operable to anchor the composition to the location using an adhesive, negative pressure and/or fastener. In some embodiments of any of the methods described herein, the reservoir is an anchorable reservoir.
In some embodiments of any of the methods described herein, the pharmaceutical composition is an ingestible device, that includes: a housing; a reservoir located within the housing and containing the immune modulator, a mechanism for releasing the immune modulator from the reservoir; and an exit valve configured to allow the immune modulator to be released out of the housing from the reservoir. In some embodiments of any of the methods described herein, the ingestible device further includes: an electronic component located within the housing; and a gas generating cell located within the housing and adjacent to the electronic component, where the electronic component is configured to activate the gas generating cell to generate gas. In some embodiments of any of the methods described herein, the ingestible device further includes: a safety device placed within or attached to the housing, where the safety device is configured to relieve an internal pressure within the housing when the internal pressure exceeds a threshold level.
In some embodiments of any of the methods described herein, the pharmaceutical composition is an ingestible device, that includes: a housing defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end; an electronic component located within the housing; a gas generating cell located within the housing and adjacent to the electronic component, where the electronic component is configured to activate the gas generating cell to generate gas; a reservoir located within the housing, where the reservoir stores a dispensable substance and a first end of the reservoir is attached to the first end of the housing; an exit valve located at the first end of the housing, where the exit valve is configured to allow the dispensable substance to be released out of the first end of the housing from the reservoir; and a safety device placed within or attached to the housing, where the safety device is configured to relieve an internal pressure within the housing when the internal pressure exceeds a threshold level.
In some embodiments of any of the methods described herein, the pharmaceutical composition is an ingestible device, that includes: a housing defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end; an electronic component located within the housing, a gas generating cell located within the housing and adjacent to the electronic component, where the electronic component is configured to activate the gas generating cell to generate gas; a reservoir located within the housing, where the reservoir stores a dispensable substance and a first end of the reservoir is attached to the first end of the housing; an injection device located at the first end of the housing, where the jet injection device is configured to inject the dispensable substance out of the housing from the reservoir; and a safety device placed within or attached to the housing, where the safety device is configured to relieve an internal pressure within the housing.
In some embodiments of any of the methods described herein, the pharmaceutical composition is an ingestible device, that includes: a housing defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end; an optical sensing unit located on a side of the housing, where the optical sensing unit is configured to detect a reflectance from an environment external to the housing; an electronic component located within the housing; a gas generating cell located within the housing and adjacent to the electronic component, where the electronic component is configured to activate the gas generating cell to generate gas in response to identifying a location of the ingestible device based on the reflectance; a reservoir located within the housing, where the reservoir stores a dispensable substance and a first end of the reservoir is attached to the first end of the housing; a membrane in contact with the gas generating cell and configured to move or deform into the reservoir by a pressure generated by the gas generating cell; and a dispensing outlet placed at the first end of the housing, where the dispensing outlet is configured to deliver the dispensable substance out of the housing from the reservoir.
In some embodiments, provided herein is a method of treating a disease as disclosed herein, comprising:
administering to the subject a pharmaceutical formulation that comprises a therapeutic agent as disclosed herein, wherein the pharmaceutical formulation is released at a location in the gastrointestinal tract of the subject, such as a location that is proximate to one or more sites of disease.
In some embodiments, the pharmaceutical formulation is administered in an ingestible device. In some embodiments, the pharmaceutical formulation is released from an ingestible device. In some embodiments, the ingestible device comprises a housing, a reservoir containing the pharmaceutical formulation, and a release mechanism for releasing the pharmaceutical formulation from the device, wherein the reservoir is releasably or permanently attached to the exterior of the housing or internal to the housing.
In some embodiments, provided herein is a method of treating a disease as disclosed herein, comprising:
administering to the subject an ingestible device comprising a housing, a reservoir containing a pharmaceutical formulation, and a release mechanism for releasing the pharmaceutical formulation from the device, wherein the reservoir is releasably or permanently attached to the exterior of the housing or internal to the housing;
wherein the pharmaceutical formulation comprises a therapeutic agent as disclosed herein, and the ingestible device releases the pharmaceutical formulation at a location in the gastrointestinal tract of the subject, such as a location that is proximate to one or more sites of disease.
In some embodiments, the housing is non-biodegradable in the GI tract.
In some embodiments, the release of the formulation is triggered autonomously.
In some embodiments, the device is programmed to release the formulation with one or more release profiles that may be the same or different at one or more locations. In some embodiments, the device is programmed to release the formulation at a location proximate to one or more sites of disease. In some embodiments, the location of one or more sites of disease is predetermined.
In some embodiments, the reservoir is made of a material that allows the formulation to leave the reservoir, such as a biodegradable material.
In some embodiments, the release of the formulation is triggered by a pre-programmed algorithm. In some embodiments, the release of the formulation is triggered by data from a sensor or detector to identify the location of the device. In some more particular embodiments, the data is not based solely on a physiological parameter (such as pH, temperature, and/or transit time).
In some embodiments, the device comprises a detector configured to detect light reflectance from an environment external to the housing. In some more particular embodiments, the release is triggered autonomously or based on the detected reflectance.
In some embodiments, the device releases the formulation at substantially the same time as one or more sites of disease are detected. In some embodiments, the one or more sites of disease are detected by the device (e.g., by imaging the GI tract).
In some embodiments, the release mechanism is an actuation system. In some embodiments, the release mechanism is a chemical actuation system. In some embodiments, the release mechanism is a mechanical actuation system. In some embodiments, the release mechanism is an electrical actuation system. In some embodiments, the actuation system comprises a pump and releasing the formulation comprises pumping the formulation out of the reservoir. In some embodiments, the actuation system comprises a gas generating cell.
In some embodiments, the device further comprises an anchoring mechanism.In some embodiments, the formulation comprises a therapeutically effective amount of the therapeutic agent as disclosed herein. In some embodiments, the formulation comprises a human equivalent dose (HED) of the therapeutic agent as disclosed herein.
In some embodiments, the device is a device capable of releasing a solid therapeutic agent as disclosed herein or a solid formulation comprising the therapeutic agent as disclosed herein. In some embodiments, the device is a device capable of releasing a liquid therapeutic agent as disclosed herein or a liquid formulation comprising the therapeutic agent as disclosed herein. Accordingly, in some embodiments of the methods herein, the pharmaceutical formulation release from the device is a solid formulation.
Accordingly, in some embodiments of the methods herein, the pharmaceutical formulation release from the device is a liquid formulation.
The devices disclosed herein are capable of releasing a therapeutic agent as disclosed herein or a formulation comprising the therapeutic agent as disclosed herein irrespective of the particular type of therapeutic agent as disclosed herein. For example, the therapeutic agent as disclosed herein may be a small molecule, a biological, a nucleic acid, an antibody, a fusion protein, and so on.

In some embodiments, provided herein is a method of releasing a therapeutic agent as disclosed herein into the gastrointestinal tract of a subject for treating one or more sites of disease within the gastrointestinal tract, the method comprising:
administering to the subject a therapeutically effective amount of the therapeutic agent as disclosed herein housed in an ingestible device, wherein the ingestible device comprises a detector configured to detect the presence of the one or more sites of disease, and a controller or processor configured to trigger the release of the therapeutic agent as disclosed herein proximate to the one or more sites of disease in response to the detector detecting the presence of the one or more sites of disease.
In some embodiments, provided herein is a method of releasing a therapeutic agent as disclosed herein into the gastrointestinal tract of a subject for treating one or more pre-determined sites of disease within the gastrointestinal tract, the method comprising:
administering to the subject a therapeutically effective amount of the therapeutic agent as disclosed herein contained in an ingestible device, wherein the ingestible device comprises a detector configured to detect the location of the device within the gastrointestinal tract, and a controller or processor configured to trigger the release of the therapeutic agent as disclosed herein proximate to the one or more predetermined sites of disease in response to the detector detecting a location of the device that corresponds to the location of the one or more pre-determined sites of disease.
In some embodiments, provided herein is a method of releasing a therapeutic agent as disclosed herein into the gastrointestinal tract of a subject for treating one or more sites of disease within the gastrointestinal tract, the method comprising:
administering to the subject a therapeutically effective amount of the therapeutic agent as disclosed herein contained in an ingestible device;
receiving at an external receiver from the device a signal transmitting environmental data;
assessing the environmental data to confirm the presence of the one or more sites of disease; and when the presence of the one or more sites of disease is confirmed, sending from an external transmitter to the device a signal triggering the release of the therapeutic agent as disclosed herein proximate to the one or more sites of disease.

In some embodiments, provided herein is a method of releasing a therapeutic agent as disclosed herein into the gastrointestinal tract of a subject for treating one or more sites of disease within the gastrointestinal tract, the method comprising:
administering to the subject a therapeutically effective amount of the therapeutic agent as disclosed herein contained in an ingestible device;
receiving at an external receiver from the device a signal transmitting environmental or optical data;
assessing the environmental or optical data to confirm the location of the device within the gastrointestinal tract; and when the location of the device is confirmed, sending from an external transmitter to the device a signal triggering the release of the therapeutic agent as disclosed herein proximate to the one or more sites of disease.
In some embodiments of any of the methods described herein, the pharmaceutical composition is an ingestible device as disclosed in US Patent Application Ser.
No.
62/385,553, incorporated by reference herein in its entirety. In some embodiments of any of the methods described herein, the pharmaceutical composition is an ingestible device that includes a localization mechanism as disclosed in international patent application PCT/US2015/052500, incorporated by reference herein in its entirety. In some embodiments of any of the methods described herein, the pharmaceutical composition is not a dart-like dosage form.
Also provided herein are methods of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm of a subject, that include: releasing an immune modulator at a location in the large intestine of the subject, wher the method includes administering endoscopically to the subject a therapeutically effective amount of the immune modulator, where the method does not include releasing more than 20% of the immune modulator at a location that is not an intended site of release.
Also provided herein are methods of treating a disease or condition that arises in a tissue originating from the endoderm in a subject, that include: releasing an immune modulator at a location in the proximal portion of the large intestine of the subject, where the method includes administering endoscopically to the subject a pharmaceutical composition including a therapeutically effective amount of the immune modulator, where the pharmaceutical composition is an ingestible device.

In some embodiments of any of the methods described herein, the method does not include releasing more than 20% of the immune modulator at a location that is not proximate to an intended site of release. In some embodiments of any of the methods described herein, the method does not include releasing more than 10% of the immune modulator at a location that is not proximate to an intended site of release. In some embodiments of any of the methods described herein, the method provides a concentration of the immune modulator at a location that is an intended site of release that is 2-100 times greater than at a location that is not the intended site of release. In some embodiments of any of the methods described herein, the method provides a concentration of the immune modulator in the plasma of the subject that is less than 3 [tg/mL. In some embodiments of any of the methods described herein, the method provides a concentration of the immune modulator in the plasma of the subject that is less than 0.3 [tg/mL. In some embodiments of any of the methods described herein, the method provides a concentration of the immune modulator in the plasma of the subject that is less than 0.01 [tg/mL. In some embodiments of any of the methods described herein, the method provides a C24 value of the immune modulator in the plasma of the subject that is less than 3 [tg/mL. In some embodiments of any of the methods described herein, the method provides a C24 value of the immune modulator in the plasma of the subject that is less than 0.3 g/mL. In some embodiments of any of the methods described herein, the method provides a C24 value of the immune modulator in the plasma of the subject that is less than 0.01 g/mL.
In some embodiments of any of the methods described herein, the composition does not include an enteric coating. In some embodiments of any of the methods described herein, the immune modulator is not a cyclic peptide. In some embodiments of any of the methods described herein, the immune modulator is present in a pharmaceutical formulation within the device. In some embodiments of any of the methods described herein, the formulation is a solution of the immune modulator in a liquid medium. In some embodiments of any of the methods described herein, the formulation is a suspension of the immune modulator in a liquid medium.
In some embodiments of any of the methods described herein, the tissue originating from the endoderm is selected from the group of: the stomach, the colon, the liver, the pancreas, the urinary bladder, the epithelial parts of the trachea, the lungs, the pharynx, the thyroid, the parathyroid, the intestines, and the gallbladder. In some embodiments of any of the methods described herein, the inflammatory disease or condition that arises in a tissue originating from the endoderm is selected from the group of: gastritis, Celiac disease, hepatitis, alcoholic lever disease, fatty liver disease (hepatic steatosis), non-alcoholic fatty liver disease (NASH), cirrhosis, primary schlerosing cholangitis, pancreatitis, insterstitial cystitits, asthma, chronic obstructic pulmonary disease, pulmonary fibrosis, pharyngitis, thyroiditis, hyperthyroidism, parathyroiditis, nephritis, Hashimoto's disease, Addison's disease, Graves' disease, Sjogren syndrome, type 1 diabetes, pelvic inflammatory disease, auditory canal inflammation, tinnitus, vestibular neuritis, otitis media, auditory canal inflammation, tracheitis, cholestatic liver disease, primary biliary schlerosis, liver parenchyma, an inherited metabolic disorder of the liver, Byler syndrome, cerebrotendinous, xanthomatosis, Zellweger's syndrome, neonatal hepatitis, cystic fibrosis, ALGS
(Alagilles syndrome), PFIC (progressive familial intrahepatic cholestasis), autoimmune hepatitis, primary biliary cirrhosis (PBC), liver fibrosis, NAFLD, portal hypertension, general cholestasis, such as in jaundice due to drugs or during pregnancy, intra- and extrahepatic cholestasis, such as hereditary forms of cholestasis, such as PFIC1, gall stones and choledocholithiasis, malignancy causing obstruction of the biliary tree, symptoms (scratching, pruritus) due to cholestasis/jaundice, chronic autoimmune liver disease leading to progressive cholestasis, and pruritus of cholestatic liver disease, duodenal ulcers, enteritis (radition-, chemotherapy-, or infection-induced enteritis), diverticulitis, pouchitis, cholecystitis, and cholangitis. In some embodiments of any of the methods described herein, the inflammatory disease or condition that arises in a tissue originating from the endoderm is inflammation of the liver.
In some embodiments of any of the methods described herein, the immune modulator is released at a location in the proximal portion of the ascending colon. In some embodiments of any of the methods described herein, the immune modulator is released at a location in the proximal portion of the cecum. In some embodiments of any of the methods described herein, the immune modulator is released at a location in the proximal portion of the sigmoid colon. In some embodiments of any of the methods described herein, the immune modulator is released at a location in the proximal portion of the transverse colon.
In some embodiments of any of the methods described herein, the immune modulator is released at a location in the proximal portion of the descending colon. In some embodiments of any of the methods described herein, the method includes administering to the subject a reservoir including the therapeutically effective amount of the immune modulator, where the reservoir is connected to the endoscope.

Some embodiments of any of the methods described herein further include administering a second agent orally, intravenously or subcutaneously, where the second agent is the same immune modulator; a different immune modulator; or an agent having a different biological target from the immune modulator, where the second agent is an agent suitable for treating an inflammatory disease or condition that arises in a tissue originating from the endoderm. In some embodiments of any of the methods described herein, the immune modulator is administered prior to the second agent. In some embodiments of any of the methods described herein, the immune modulator is administered after the second agent. In some embodiments of any of the methods described herein, the immune modulator and the second agent are administered substantially at the same time. In some embodiments of any of the methods described herein, the second agent is administered intravenously.
In some embodiments of any of the methods described herein, the second agent is administered subcutaneously. In some embodiments of any of the methods described herein, the amount of the second agent is less than the amount of the second agent when the immune modulator and the second agent are both administered systemically. In some embodiments of any of the methods described herein, the second agent is another immune modulator. In some embodiments of any of the methods described herein, the method does not include administering a second agent.
In some embodiments of any of the methods described herein, the method includes identifying an intended site of release prior to endoscopic administration. In some embodiments of any of the methods described herein, the method includes identifying an intended site of release substantially at the same time as releasing the immune modulator. In some embodiments of any of the methods described herein, the method includes monitoring the progress of the disease. In some embodiments of any of the methods described herein, the method does not include administering an immune modulator with a spray catheter. In some embodiments of any of the methods described herein, the method includes administering an immune modulator with a spray catheter.
Also provided herein are methods of treating an inflammatory disease or condition that arises in a tissue arising from the endoderm in a subject, that include:
releasing an immune modulator at a location in the gastrointestinal tract of the subject that is proximate to an intended site of release, where the methods include administering to the subject a pharmaceutical composition including a therapeutically effective amount of the immune modulator the method including one or more of the following steps: (a) identifying a subject having a disease or condition that arises in a tissue originating from the endoderm;
(b) determination of the severity of the disease; (c) determination of the location of the disease; (d) evaluating the subject for suitability to treatment; (e) administration of an induction dose of the immune modulator; (f) monitoring the progress of the disease; and/or (g) optionally repeating steps (e) and (f) one or more times.
In some embodiments of any of the methods described herein, the pharmaceutical composition is an ingestible device and the method includes administering orally to the subject the pharmaceutical composition. In some embodiments of any of the methods described herein, the method includes administering one or more maintenance doses following administration of the induction dose in step (e). In some embodiments of any of the methods described herein, the induction dose is a dose of the immune modulator administered in an ingestible device. In some embodiments of any of the methods described herein, the maintenance dose is a dose of the immune modulator administered in an ingestible device as disclosed herein. In some embodiments of any of the methods described herein, the maintenance dose is a dose of the immune modulator delivered systemically. In some embodiments of any of the methods described herein, the induction dose is a dose of the immune modulator delivered systemically. In some embodiments of any of the methods described herein, the maintenance dose is a dose of the immune modulator administered in an ingestible device. In some embodiments of any of the methods described herein, the induction dose is a dose of a second agent as delivered systemically. In some embodiments of any of the methods described herein, the maintenance dose is a dose of the immune modulator administered in an ingestible device.
In some embodiments of any of the methods described herein, wherein the immune modulator is selected from the group of: IL-12/IL-23 inhibitors, TNFa inhibitors, IL-6 receptor inhibitors, CD40/CD4OL inhibitors, IL-1 inhibitors, IL-13 inhibitors, IL-10 receptor agonists, and integrin inhibitors. In some embodiments of any of the methods described herein, the immune modulator is a PDE4 inhibitor.
Also provided herein are immune modulator delivery apparatuses that include:
an ingestible housing including a reservoir having a pharmaceutical composition including a therapeutically effective amount of the immune modulator stored therein; a detector coupled to the ingestible housing, the detector configured to detect when the ingestible housing is proximate to a respective intended site of release; a valve system in fluid communication with the reservoir system; and a controller communicably coupled to the valve system and the detector, the controller configured to cause the valve system to open in response to the detector detecting that the ingestible housing is proximate to the respective intended site of release so as to release the therapeutically effective amount of the immune modulator at the respective intended site of release. Some embodiments of any of the apparatuses described herein further include a pump positioned in the ingestible housing, the pump configured to pump the therapeutically effective amount of the immune modulator from the reservoir in response to activation of the pump by the controller responsive to detection by the detector of the ingestible housing being proximate to the intended site of release. In some embodiments .. of any of the apparatuses described herein, the controller is configured to cause the pump to pump the therapeutically effective amount of the immune modulator from the reservoir according to the following protocol. In some embodiments of any of the apparatuses described herein, the valve system includes a dissolvable coating. In some embodiments of any of the apparatuses described herein, the valve system includes one or more doors configured for actuation by at least one of sliding, pivoting, and rotating.
In some embodiments of any of the apparatuses described herein, the valve system includes an electrostatic shield. In some embodiments of any of the apparatuses described herein, the reservoir includes a pressurized cell.
Some embodiments of any of the apparatuses described herein further include at least one actuatable anchor configured to retain the ingestible housing at the respective intended site of release upon actuation. In some embodiments of any of the apparatuses described herein, the actuatable anchor is retractable.
Also provided herein are compositions that include a therapeutically effective amount of any of the immune modulators described herein, where the composition is capable of releasing the immune modulator at a location in the gastrointestinal tract of the subject. In some embodiments of any of the compositions described herein, the composition includes a tissue anchoring mechanism for anchoring the composition to the location. In some embodiments of any of the compositions described herein, the tissue anchoring mechanism is capable of anchoring for anchoring to the location. In some embodiments of any of the compositions described herein, the tissue anchoring mechanism includes an osmotically-driven sucker. In some embodiments of any of the compositions described herein, the tissue anchoring mechanism comprises a connector operable to anchor the composition to the location. In some embodiments of any of the compositions described herein, the connector is operable to anchor the composition to the location using an adhesive, negative pressure and/or fastener.
Also provided herein is an immune modulator for use in a method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm in a subject, where the method includes orally administering to the subject an ingestible device loaded with the immune modulator, wherein the immune modulator is released by the device at a location in the gastrointestinal tract of the subject that is proximate to an intended site of release of the immune modulator. In some embodiments of an immune modulator for use described herein, the immune modulator is contained in a reservoir suitable for attachment to .. a device housing, and wherein the method includes attaching the reservoir to the device housing to form the ingestible device, prior to orally administering the ingestible device to the subject.
Also provided herein is an attachable reservoir containing an immune modulator for use in a method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm, where the method includes attaching the reservoir to a device housing to form an ingestible device and orally administering the ingestible device to a subject, where the immune modulator is released by device at a location in the gastrointestinal tract of the subj ect that is proximate to the intended site of release.
Also provided herein is a composition including or consisting of an ingestible device .. loaded with a therapeutically effective amount of an immune modulator, for use in a method of treatment, wherein the method includes orally administering the composition to the subject, wherein the immune modulator is released by the device at a location in the gastrointestinal tract of the subject that is proximate to an intended site of release.
In some embodiments of any of the immune modulators for use described herein, any .. of the attachable reservoirs described herein, or the compositions for use described herein, the intended site of release has been pre-determined. In some embodiments of any of the immune modulators for use described herein, any of the attachable reservoirs described herein, or any of the compositions for use described herein, the ingestible device further includes an environmental sensor and the method further includes using the environmental sensor to identify the location of the intended site of release. In some embodiments of any of the immune modulators for use, any of the attachable reservoirs described herein, or any of the compositions for use described herein, the environmental sensor is an imaging sensor and the method further includes imaging the gastrointestinal tract to identify the intended site of release. In some embodiments of any of the immune modulators for use described herein, any of the attachable reservoirs described herein, or any of the compositions for use described herein, the imaging detects an intended site of release. In some embodiments of any of the immune modulators for use, any of the attachable reservoirs described herein, or any of the compositions for use described herein, the inflammatory disease or condition that arises in a tissue originating from the endoderm is selected from the group of: gastritis, Celiac disease, hepatitis, alcoholic liver disease, fatty liver disease (hepatic steatosis), non-alcoholic fatty liver disease (NASH), cirrhosis, primary schlerosing cholangitis, pancreatitis, insterstitial cystitits, asthma, chronic obstructic pulmonary disease, pulmonary fibrosis, pharyngitis, .. thyroiditis, hyperthyroidism, parathyroiditis, nephritis, Hashimoto's disease, Addison's disease, Graves' disease, Sjogren syndrome, type 1 diabetes, pelvic inflammatory disease, auditory canal inflammation, tinnitus, vestibular neuritis, otitis media, auditory canal inflammation, tracheitis, cholestatic liver disease, primary biliary schlerosis, liver parenchyma, an inherited metabolic disorder of the liver, Byler syndrome, cerebrotendinous, xanthomatosis, Zellweger's syndrome, neonatal hepatitis, cystic fibrosis, ALGS
(Alagilles syndrome), PFIC (progressive familial intrahepatic cholestasis), autoimmune hepatitis, primary biliary cirrhosis (PBC), liver fibrosis, NAFLD, portal hypertension, general cholestasis, such as in jaundice due to drugs or during pregnancy, intra- and extrahepatic cholestasis, such as hereditary forms of cholestasis, such as PFIC1, gall stones and .. choledocholithiasis, malignancy causing obstruction of the biliary tree, symptoms (scratching, pruritus) due to cholestasis/jaundice, chronic autoimmune liver disease leading to progressive cholestasis, and pruritus of cholestatic liver disease, duodenal ulcers, enteritis (radition-, chemotherapy-, or infection-induced enteritis), diverticulitis, pouchitis, cholecystitis, and cholangitis.
In some embodiments of any of the immune modulators for use, any of the attachable reservoirs described herein, or any of the compositions for use described herein, the inflammatory disease or condition that arises in a tissue originating from the endoderm is a liver disease or disorder selected from the group of: fibrosis, cirrhosis, alcoholic lever disease, fatty liver disease (hepatic steatosis), non-alcoholic fatty liver disease (NASH), cholestatic liver disease, liver parenchyma, an inherited metabolic disorder of the liver, PFIC
(progressive familial intrahepatic cholestasis), autoimmune hepatitis, primary biliary cirrhosis (PBC), NAFLD, chronic autoimmune liver disease leading to progressive cholestasis, pruritus of cholestatic liver disease, inflammation of the liver, and liver fibrosis.

In some embodiments of any of the immune modulators for use, any of the attachable reservoirs described herein, or any of the compositions for use described herein, the disease or condition that arises in a tissue originating from the endoderm is a disease or condition related to the gut-brain axis selected from the group consisting of multiple sclerosis, Parkinson's disease, mild cognitive impairment, Alzheimer's, disease, encephalitis, and hepatic encephalopathy.
Also provided herein are ingestible devices loaded with a therapeutically effective amount of an immune modulator, where the device is controllable to release the immune modulator at a location in the gastrointestinal tract of the subject that is proximate to an intended site of release. Also provided herein are any of the devices described herein for use in a method of treatment of the human or animal body.
In some embodiments of any of the immune modulators for use described herein, any of the attachable reservoirs described herein, or any of the devices described herein, wherein the ingestible device includes: a housing defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end; a reservoir located within the housing and containing the immune modulator, where a first end of the reservoir is connected to the first end of the housing; a mechanism for releasing the immune modulator from the reservoir; and an exit value configured to allow the immune modulator to be released out of the housing from the reservoir.
In some embodiments of any of the immune modulators for use described herein, any of the attachable reservoirs described herein, or any of the devices described herein, the ingestible device includes: an ingestible housing including a reservoir compartment having a therapeutically effective amount of the immune modulator stored therein; a release mechanism having a closed state which retains the immune modulator in the reservoir and an open state which releases the immune modulator the reservoir to the exterior of the device;
and an actuator which changes the state of the release mechanism from the closed to the open state.
In some embodiments of any of the immune modulators for use described herein, any of the attachable reservoirs described herein, or any of the devices described herein, the .. ingestible device further comprises an environmental sensor for detecting the location of the device in the gut. In some embodiments of any of the immune modulators for use described herein, any of the compositions for use described herein, or any of the devices described herein, where the ingestible device further includes a communication system for transmitting data from the environmental sensor to an external receiver. In some embodiments of any of the immune modulators for use described herein, any of the attachable reservoirs described herein, any of the compositions for use described herein, or any of the devices described herein, the ingestible device further includes a processor or controller which is coupled to the environmental sensor and to the actuator and which triggers the actuator to cause the release mechanism to transition from its closed state to its open state when it is determined that the device is in the presence of the intended site of release and/or is in a location in the gut that has been predetermined to be proximal to the intended site of release.
In some embodiments of any of the immune modulators for use described herein, any of the attachable reservoirs described herein, any of the compositions for use described herein, or any of the devices described herein, the communication system further includes means for receiving a signal from an external transmitter, and where the actuator is adapted to be triggered in response to the signal.
In some embodiments of any of the immune modulators for use described herein, any of the attachable reservoirs described herein, any of the compositions for use described herein, or any of the devices described herein, the ingestible device further includes a communication system for transmitting localization data to an external receiver.
In some embodiments of any of the immune modulators for use described herein, any of the attachable reservoirs described herein, any of the compositions for use described .. herein, or any of the devices described herein, the ingestible device further includes a communication system for transmitting localization data to an external receiver and for receiving a signal from an external transmitter; where the actuator is adapted to be triggered in response to the signal. In some embodiments of any of the immune modulators for use described herein, any of the attachable reservoir compartments for use described herein, any of the compositions for use described herein, or any of the devices described herein, the ingestible device further includes a deployable anchoring system and an actuator for deploying the anchoring system, where the anchoring system is capable of anchoring or attaching the ingestible device to the subject's tissue.
In some embodiments of any of the methods described herein, the subject has previously been identified as having an inflammatory disease or condition that arises in a tissue originating from the endoderm.
Aspects and embodiments as described herein are intended to be freely combinable.
For example, any details or embodiments described herein for methods of treatment apply equally to an agent, composition or ingestible device for use in said treatment. Any details or embodiments described for a device apply equally to methods of treatment using the device, or to an agent or composition for use in a method of treatment involving the device.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is a view of an example embodiment of an ingestible device, in accordance with some embodiments of the disclosure.
FIG. 2 is an exploded view of the ingestible device of FIG. 1, in accordance with some embodiments of the disclosure.
FIG. 3 is a diagram of an ingestible device during an example transit through a GI
tract, in accordance with some embodiments of the disclosure.
FIG. 4 is a diagram of an ingestible device during an example transit through a jejunum, in accordance with some embodiments of the disclosure.
FIG. 5 is a flowchart of illustrative steps for determining a location of an ingestible .. device as it transits through a GI tract, in accordance with some embodiments of the disclosure.
FIG. 6 is a flowchart of illustrative steps for detecting transitions from a stomach to a duodenum and from a duodenum back to a stomach, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.
FIG. 7 is a plot illustrating data collected during an example operation of an ingestible device, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.
FIG. 8 is another plot illustrating data collected during an example operation of an ingestible device, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.
FIG. 9 is a flowchart of illustrative steps for detecting a transition from a duodenum to a jejunum, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.
FIG. 10 is a plot illustrating data collected during an example operation of an ingestible device, which may be used when detecting a transition from a duodenum to a jejunum, in accordance with some embodiments of the disclosure.

FIG. 11 is a plot illustrating muscle contractions detected by an ingestible device over time, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.
FIG. 12 is a flowchart of illustrative steps for detecting a transition from a jejunum to an ileum, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.
FIG. 13 is a flowchart of illustrative steps for detecting a transition from a jejunum to an ileum, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.
FIG. 14 is a flowchart of illustrative steps for detecting a transition from an ileum to a cecum, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.
FIG. 15 is a flowchart of illustrative steps for detecting a transition from a cecum to a colon, which may be used when determining a location of an ingestible device as it transits through a GI tract, in accordance with some embodiments of the disclosure.
FIG. 16 illustrates an ingestible device for delivering a substance in the GI
tract.
FIG. 17 illustrates aspects of a mechanism for an ingestible device with a gas generating cell configured to generate a gas to dispense a substance.
FIG. 18 illustrates an ingestible device having a piston to push for drug delivery.
FIG. 19 illustrates an ingestible device having a bellow structure for a storage reservoir of dispensable substances.
FIG. 20 illustrates an ingestible device having a flexible diaphragm to deform for drug delivery.
FIG. 21 shows an illustrative embodiment of an ingestible device with multiple openings in the housing.
FIG. 22 shows a highly cross-section of an ingestible device including a valve system and a sampling system.
FIG. 23 illustrates a valve system.
FIGs. 24A and 24B illustrate a portion of a two-stage valve system in its first and second stages, respectively.
FIGs. 25A and 25B illustrate a portion of a two-stage valve system in its first and second stages, respectively.

FIGs. 26A and 26B illustrate a portion of a two-stage valve system in its first and second stages, respectively.
FIG. 27 illustrates a more detailed view of an ingestible device including a valve system and a sampling system.
FIG. 28 illustrates a portion of an ingestible device including a sampling system and a two-stage valve system in its second stage.
FIG. 29 is a highly schematic illustrate of an ingestible device.
FIG. 30 is a graph shiwng the percentage (%) change in body weight at day 14 ( SEM) for DSS mice treated with anti-IL-12 p40 antibody intraperitoneally (10 mg/kg) every third day (Q3D) or intracecally (10 mg/kg or 1 mg/kg) daily (QD), when compared to mice treated with anti-IL-12 p40 antibody intraperitoneally (10 mg/kg) every third day (Q3D) and vehicle control (Vehicle). Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p <0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 31 is a graph showing the concentration of anti-IL-12 p40 rat IgG2A ( g/mL) in plasma of anti-IL-12 p40 intraperitoneally (10 mg/kg) and intracecally (10 mg/kg and 1 mg/kg) administered treatment groups given daily (QD) or every third day (Q3D) when compared to vehicle control (Vehicle) and when IP is compared to IC. ELISA
analysis was used to determine the concentration of anti-IL-12 p40 (IgG2A). Data presented as mean SEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p <0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 32 is a graph showing the concentration of anti-IL-12 p40 antibody (IgG2A) ( g/mL) in the cecum and colon content of anti-IL-12 p40 antibody intraperitoneally (10 mg/kg) and intracecally (10 mg/kg and 1 mg/kg) administered treatment groups given daily (QD) or every third day (Q3D), when compared to vehicle control (Vehicle) and when IP is compared to IC. ELISA analysis was used to determine the concentration of rat IgG2A. Data presented as mean SEM. Mann-Whitney's U- test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p < 0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 33 is a graph showing the mean overall tissue immunolabel scores (intensity and extent) in acute DSS colitis mouse colon of anti-IL-12 p40 antibody intracecally-treated versus vehicle control-treated DSS mice. Data presented as mean SEM.

FIG. 34 is a graph showing the mean location-specific immunolabel scores in acute DSS colitis mouse colon of anti-IL-12 p40 intracecally-treated versus vehicle control-treated DSS mice. Data presented as mean SEM. Mann-Whitney's U- test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p < 0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 35 is a graph showing the ratio of anti-IL-12 p40 antibody in the colon tissue to the plasma concentration of the anti-IL-12 p40 antibody in mice treated with the anti-IL-12 p40 antibody on day 0 (QO) or day 3 (Q3D) of the study, when measured at the same time point after the initial dosing. An outlier animal was removed from Group 5.
FIG. 36 is a graph showing the concentration of I1-113 ( g/mL) in colon tissue lysate of acute DSS colitis mice treated with anti-IL-12 p40 intraperitoneally (10 mg/kg) every third day (Q3D) or intracecally (10 mg/kg or 1 mg/kg) adminitsered daily (QD), when compared to vehicle control (Vehicle). Data presented as mean SEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data .. respectively. A value of p < 0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 37 is a graph showing the concentration of 11-6 ( g/mL) in colon tissue lysate of acute DSS colitis mice treated with anti-IL-12 p40 intraperitoneally (10 mg/kg) every third day (Q3D) or intracecally (10 mg/kg or 1 mg/kg) administered daily (QD), when compared to vehicle control (Vehicle). Data presented as mean SEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p < 0.05 was considered significant (Graph Pad Software, Inc.
FIG. 38 is a graph showing the concentration of I1-17A ( g/mL) in colon tissue lysate of acute DSS colitis mice treated with anti-IL-12 p40 intraperitoneally (10 mg/kg) every third day (Q3D) or intracecally (10 mg/kg and 1 mg/kg) administered daily (QD), when compared to vehicle control (Vehicle). Data presented as mean SEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p < 0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 39 is a graph showing the percentage (%) change in body weight at day 14 ( SEM) for DSS mice treated with DATK32 (anti-a4137) antibody intraperitoneally (25 mg/kg) every third day (Q3D) or intracecally (25 mg/kg or 5 mg/kg) administered daily (QD), when compared to vehicle control (Vehicle) and when IC is compared to IP. Data presented as mean SEM. Mann-Whitney's U- test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value ofp < 0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 40 is a graph showing the plasma concentration of DATK32 rat IgG2A ( g/mL) of intraperitoneally (25mg/kg) and intracecally (25 mg/kg and 5 mg/kg) administered treatment groups given daily (QD) or every third day (Q3D), where IP is compared to IC.
Data presented as mean SEM. Mann-Whitney's U- test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p < 0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 41 is a graph showing the concentration of DATK32 rat IgG2A antibody ( g/mL) in cecum and colon content of intraperitoneally (25mg/kg) or intracecally (25 mg/kg and 5 mg/kg) administered treatment groups given daily (QD) or every third day (Q3D), where IP is compared to IC. Data presented as mean SEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p < 0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 42 is a graph showing the concentration of DATK32 rat IgG2A ( g/mL) in the colon content of intraperitoneally (25mg/kg) or intracecally (25 mg/kg and 5 mg/kg) administered treatment groups given daily (QD), and concentration over time (1, 2 ,4, 24, and 48 hours), where IP is compared to IC. Data presented as mean SEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value ofp<0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 43 is a graph showing the concentration of DATK32 rat IgG2A ( g/g) in colon tissue of intraperitoneally (25mg/kg) or intracecally (25 mg/kg and 5 mg/kg) administered treatment groups given daily (QD) or every third day (Q3D), where IP is compared to IC.
Data presented as mean SEM. Mann-Whitney's U- test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value ofp<0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 44 is a graph showing the concentration of DATK32 rat IgG2A ( g/g) in the colon tissue of intraperitoneally (25mg/kg) or intracecally (25 mg/kg and 5 mg/kg) administered treatment groups given daily (QD), and the concentration over time (1, 2, 4, 24, and 48 hours) was determined, where IP is compared to IC. Data presented as mean SEM.
Mann-Whitney's U- test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p < 0.05 was considered significant (Graph Pad Software, Inc.).

FIG. 45 is a graph showing the mean overall tissue immunolabel scores (intensity and extent) in acute DSS colitis mouse colon of DATK32 (anti-a4137) antibody treated versus vehicle control (Vehicle) treated DSS mice. The data are presented as mean SEM.
FIG. 46 is a graph showing the mean location-specific immunolabel scores in acute DSS colitis mouse colon of DATK32 (anti-a4137) antibody-treated versus vehicle control (Vehicle)-treated DSS mice. Data presented as mean SEM. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value ofp < 0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 47 is a graph showing the ratio of the DATK-32 antibody in the colon tissue to the plasma concentration of the DATK-32 antibody in mice treated with the DATK-antibody on day 0 (QO) or day 3 (Q3D) of the study (Groups 9-12), when measured after initial dosing.
FIG. 48 is a graph showing the mean percentage of Th memory cells (mean SEM) in blood for DATK32 (anti-a4137) antibody intraperitoneally (25mg/kg) or intracecally (25 mg/kg or 5 mg/kg) administered treatment groups given daily (QD) or every third day (Q3D), when compared to vehicle control (Vehicle) and when IP is compared to IC. Mean percentage Th memory cells were measured using FACS analysis. Data presented as mean SEM. Mann-Whitney's U- test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value ofp < 0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 49 is an exemplary image of a histological section of a distal transverse colon of Animal 1501 showing no significant lesions (i.e., normal colon).
FIG. 50 is an exemplary image of a histological section of a distal transverse colon of Animal 2501 (treated with TNBS) showing areas of necrosis and inflammation.
FIG. 51 is a representative graph of plasma adalimumab concentrations over time following a single subcutaneous (SQ) or topical administration of adalimumab.
The plasma concentrations of adalimumab were determined 6, 12, 24, and 48 hours after administration of adalimumab. N/D = not detectable.
FIG. 52 is a representative table of the plasma adalimumab concentrations (1.tg/mL) as shown in FIG. 4.6.
FIG. 53 is a graph showing the concentration of TNFa (pg/mL per mg of total protein) in non-inflamed and inflamed colon tissue after intracecal administration of adalimumab, as measured 6, 12, 24, and 24 hours after the initial dosing.

FIG. 54 is a graph showing the concentration of TNFa (pg/mL per mg of total protein) in colon tissue after subcutaneous or intracecal (topical) administration of adalimumab, as measured 48 hours after the initial dosing.
FIG. 55 is a graph showing the percentage (%) change in body weight at day 14 ( SEM) in acute DSS colitis mice treated with cyclosporine A orally (10 mg/kg) every third day (Q3D) or intracecally (10 mg/kg or 3 mg/kg) daily (QD), when compared to vehicle control (Vehicle). Data presented as mean SEM. Mann-Whitney's U- test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively.
A value ofp <0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 56 is a graph showing the plasma cyclosporine A (CsA) (ng/mL) concentration over time (1 h, 2 h, 4 h, and 24 h) in acute DSS colitis mice treated daily (QD) with orally (PO) (10 mg/kg) or intracecally (IC) (10 mg/kg or 3 mg/kg) administered CsA.
Data presented as mean SEM.
FIG. 57 is a graph showing the colon tissue cyclosporine A (CsA) (ng/g) concentration over time (1 h, 2 h ,4 h and 24 h) in acute DSS colitis mice treated daily (QD) with orally (PO) (10 mg/kg) or intracecally (IC) (10 mg/kg or 3 mg/kg) administered CsA.
Data presented as mean SEM.
FIG. 58 is a graph showing the peak colon tissue cyclosporine A (CsA) (ng/g) concentration in acute DSS colitis mice treated daily (QD) with orally (PO) (10 mg/kg) or intracecally (IC) (10 mg/kg or 3 mg/kg) administered CsA. Data presented as mean SEM.
FIG. 59 is a graph showing the trough tissue concentration of cyclosporine (CsA) (ng/g) in colon of acute DSS colitis mice treated daily (QD) with orally (PO) (10 mg/kg) or intracecally (IC) (10 mg/kg or 3 mg/kg) administered CsA. Data presented as mean SEM.
FIG. 60 is a graph showing the interleukin-2 (I1-2) concentration ( g/mL) in colon tissue of acute DSS colitis mice treated daily (QD) with orally (PO) (10 mg/kg) or intracecally (IC) (10 mg/kg or 3 mg/kg) administered CsA, where PO is compared to IC.
Data presented as mean SEM. Mann-Whitney's U- test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p < 0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 61 is a graph showing the interleukin-6 (I1-6) concentration ( g/mL) in colon tissue of acute DSS colitis mice treated daily (QD) with orally (PO) (10 mg/kg) or intracecally (IC) (10 mg/kg or 3 mg/kg) administered CsA. Data presented as mean SEM.

FIG. 62 illustrates a nonlimiting example of a system for collecting, communicating and/or analyzing data about a subject, using an ingestible device.
FIGs. 63A-63F are graphs showing rat IgG2A concentration as measured in (A) colon homogenate, (B) mLN homogenate, (C) small intestine homogenate, (D) cecum contents, (E) colon contents, and (F) plasma by ELISA. Standards were prepared with plasma matrix.
Samples were diluted 1:50 before analysis. Sample 20 was removed from cecum contents analysis graph (outlier). *p<0.05; **p<0.01; ****p<0.001 were determined using the unpaired t test.
FIG. 64 illustrates a tapered silicon bellows.
FIG. 65 illustrates a tapered silicone bellows in the simulated device jig.
FIG. 66 illustrates a smooth PVC bellows.
FIG. 67 illustrates a smooth PVC bellows in the simulated device jig.
FIG. 68 demonstrates a principle of a competition assay performed in an experiment.
FIG. 69 shows AlphaLISA data.
FIG. 70 shows AlphaLISA data.
FIG. 71 shows AlphaLISA data.
FIG. 72 is a flowchart of illustrative steps of a clinical protocol, in accordance with some embodiments of the disclosure.
FIG. 73 is a graph showing the level of FAM-SMAD7-AS oligonucleotide in the cecum tissue of DSS-induced colitis mice at 12-hours. The bars represent from left to right, Groups 2 through 5 in the experiment described in Example 9.
FIG. 74 is a graph showing the level of FAM-SMAD7-AS oligonucleotide in the colon tissue of DSS-induced colitis mice at 12-hours. The bars represent from left to right, Groups 2 through 5 in the experiment described in Example 9.
FIG. 75 is a graph showing the level of FAM-SMAD7-AS oligonucleotide in the cecum contents of DSS-induced colitis mice at 12-hours. The bars represent from left to right, Groups 2 through 5 in the experiment described in Example 9.
FIG. 76 is a graph showing the mean concentration of tacrolimus in the cecum tissue and the proximal colon tissue 12 hours after intra-cecal or oral administration of tacrolimus to swine as described in Example 10.
FIG. 77 is a graph showing the mean concentration of tacrolimus in the blood 1 hour, 2 hours, 3 hours, 4 hours, 6 hours and 12 hours after intra-cecal (IC) or oral administration (PO) of tacrolimus to swine as described in Example 13.

FIG. 78 is a graph showing the AUCo-12 hours of tacrolimus in the blood after intra-cecal (IC) or oral administration (PO) of tacrolimus in swine as described in Example 13.
FIG. 79 is a graph showing the mean concentration of tacrolimus in the cecum tissue, the proximal colon tissue, the spiral colon tissue, the transverse colon tissue, and the distal colon tissue after intra-cecal (IC) or oral administration (PO) of tacrolimus in swine as described in Example 13. **** P<0.0001, *** P<0.001.
FIG. 80 is a graph showing the mean concentation of tacrolimus in the cecum lumen, the proximal lumen, the spiral colon lumen, the transverse colon lumen, and the distal colon lumen in swine after intra-cecal (IC) or oral administration (PO) of tacrolimus in swine as described in Example 13. **** P<0.0001, *** P<0.001 FIG. 81 is a bar graph showing the mean concentration of tacrolimus in the rectal content 1 hour, 3 hours, 6 hours and 12 hours after intra-cecal (IC) or oral administration (PO) of tacrolimus to swine as described in Example 13.
FIG. 82 is a line graph showing the mean concentration of tacrolimus in the rectal content 1 hour, 3 hours, 6 hours and 12 hours after intra-cecal (IC) or oral administration (PO) of tacrolimus to swine as described in Example 13.
FIG. 83 is a graph showing the mean concentration of a SMAD7 antisense molecuile (SMAD7-AS-FAM) in the cecum tissue in untreated swine or in swine after intra-cecal (IC) or oral administration(P0) of SMAD7-AS-FAM as described in Example 9.
FIG. 84 is a graph showing the mean concentration of SMAD7-AS-FAM in the colon tissue in untreated swine or in swine after intra-cecal (IC) or oral administration(P0) of SMAD7-AS-FAM as described in Example 9.
FIG. 85 is a graph showing the mean concentration of SMAD7-AS-FAM in the colon contents in untreated swine or in swine after intra-cecal (IC) or oral administration(P0) of SMAD7-AS-FAM as described in Example 9.
FIG. 86 is a graph showing the mean concentration of SMAD7-AS-FAM in the cecum contents in untreated swine or in swine after intra-cecal (IC) or oral administration(P0) of SMAD7-AS-FAM as described in Example 9.
FIG. 87 is a graph showing the mean concentration of tacrolimus in the blood of swine 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, and 12 hours after intra-cecal (IC) or oral administration (PO) of tacrolimus as described in Example 10.
FIG. 88 is a graph showing the AUCo-12 hours of tacrolimus in the blood of swine after intra-cecal (IC) or oral administration (PO) of tacrolimus as described in Example 10.

FIG. 89 is a representative table showing the Tmax, Cmax, trough (at 12 hours post-administration), and AUC0-12 hours of tacrolimus in swine after intra-cecal (IC) or oral administration (PO) as described in Example 10.
FIG. 90 is a graph showing the mean concentration of tacrolimus in the cecum, the proximal colon, the spiral colon, the transverse colon, and the distal colon of swine after intra-cecal (IC) or oral administration (PO) of tacrolimus as described in Example 10.
FIG. 91 is a graph showing the mean concentration of tacrolimus in the cecum lumen, the proximal colon lumen, the spiral colon lumen, the transverse colon lumen, and the distal colon lumen of swine after intra-cecal (IC) or oral administration (PO) of tacrolimus as described in Example 10.
FIG. 92 is a graph showing the mean concentration of tacrolimus in the rectal content of swine at 1 hour, 3 hours, 6 hours, and 12 hours after intra-cecal (IC) or oral administration (PO) of tacrolimus as described in Example 10.
FIG. 93 is a representative table showing the quantitative histological grading of colitis as described in Example 11.
FIG. 94 is a graph showing the histopathological scores of two slides for animal 1502 (healthy control swine treated with placebo), animal 2501 (swine with 8.5% DSS-induced colitis treated with 1.86 mg/kg adalimumab), animal 2503 (swine with 8.5% DSS-induced colitis treated with 1.86 mg/kg adalimumab), and animal 2504 (swine with 8.5%
DSS-induced colitis treated with 1.86 mg/kg adalimumab) at the placebo or adalimumab administration site prior to administration of placebo or adalimumab, respect tively. Absence of a bar for a particular parameter indicates that the value for this parameter was 0.
FIG. 95 is a representative hematoxylin- and eosin-stained image of the transverse colon of animal 1501 (healthy control swine). M, mucosa; SM, submucosa; TM, tunica muscularis. Numerous intestinal crypts (asterisks) are present and the surface epithelium (top two arrows) is intact. Mononuclear inflammatory cells are prominent in the lamina propria (light arrows) of the mucosa and extend a short distance into the submucosa (bottom two arrows). This amount of inflammatory cell infiltrate was expected background change and considered unrelated to the experimental protocol.
FIG. 96 is a representative hematoxylin- and eosin-stained image of the transverse colon of animal 2504 (8.5% DSS-induced colitis swine administered 1.86 mg/kg adalimumab) prior to administration of adalimumab. M, mucosa; SM, submucosa;
TM, tunica muscularis. Extensive loss (light asterisks) of intestinal crypts is present in the mucosa. Scattered crypts remain (dark asterisks) and are often dilated and filled with inflammatory cell debris and mucus. The luminal epithelium persists in some areas (upper left arrow), but is absent in others (erosion; top middle and top right arrows). Inflammatory cells in the mucosa (light arrow) are abundant and extend into the submucosa (bottom left and bottom middle arrows).
FIG. 97 is a representative immunohistochemistry micrograph of the transverse colon of animal 1501 (healthy control swine) stained for human IgG. M, mucosa; SM, submucosa;
TM, tunica muscularis. Serosal surface (arrows) and loose connective mesentery tissue (asterisks) are indicated. Faint 3,3-diaminobenzidine (DAB) staining in this tissue was considered a background effect and not indicative of human IgG.
FIG. 98 is a representative immunohistochemistry micrograph of the transverse colon of animal 2504 (8.5% DSS-induced colitis swine treated with 1.86 mg/kg dose of adsalimumab) stained for human IgG. M, mucosa; SM, submucosa; TM, tunica muscularis.
DAB staining demonstrates the presence of human IgG at the surface of luminal epithelium (two top right arrows) and at the luminal surface of an area of inflammation and erosion (top two left arrows). Intense staining is also present in the loose connective mesentery tissue (asterisks) and extends a short distance into the outer edge of the tunica muscularis (bottom left two arrows). This type of staining was considered strong (grade 4) or very strong (grade 5).
FIG. 99 is a representative immunohistochemistry micrograph of the large intestine of animal 2504 (8.5% DSS-induced colitis swine treated with 1.86 mg/kg adalimumab) stained for human IgG. M, mucosa; SM, submucosa; TM, tunica muscularis.
Lesions of DSS-induced colitis are present in this section. The luminal epithelium is absent (erosion) and diffuse loss of crypts (glands) is seen (top two asterisks). Very strong (grade 5) DAB (brown) staining demonstrates the presence of human IgG in the loose mesentery connective tissue (bottom two arterisks) and extending a short distance into the outer edge of the tunica muscularis (bottom two arrows). Strong (grade 4) staining for human IgG is seen at the eroded luminal surface (top two arrows pointing down) and within the inflammatory exudate. Weak (grade 2) staining for human IgG extends into the lamina propria (top two arrows pointing up) near the luminal surface.
FIG. 100 is a graph showing the presence of human IgG (adalimumab) at the specified locations (lumen/superficial mucosa, lamina propria, and tunica muscularis-outer/serosa) (scored level) in two slides from each of animal 1502 (placebo-treated healthy control swine), animal 2501 (swine with 8.5% DSS-induced colitis treated with 1.86 mg/kg adalimumab), animal 2503 (swine with 8.5% DSS-induced colitis treated with 1.86 mg/kg adalimumab) and animal 2504 (swine with 8.5% DSS-induced colitis treated with 1.86 mg/kg adalimumab) at the placebo or adalimumab administration site. Absence of a bar for a particular location indicates that the value for this location was 0. Scoring:
0 = not present; 1 = minimal; 2 = weak; 3 = moderate; 4 = strong; and 5=very strong immunolabel.
FIG. 101 is a graph showing the mean of Th memory cells (mean SEM) in Peyer's Patches (PP) for DATK32 antibody (anti-a4137 integrin antibody) intraperitoneally (25mg/kg) or intracecally (25 mg/kg or 5 mg/kg) administered treatment groups given daily (QD) or every third day (Q3D), when compared to vehicle control (Vehicle) and when IP
is compared to IC. Mean Th memory cells were measured using FACS analysis. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p < 0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 102 is a graph showing the mean of Th memory cells (mean SEM) in mesenteric lymph nodes (mLN) for DATK32 antibody (anti-a4137 integrin antibody) intraperitoneally (25mg/kg) or intracecally (25 mg/kg or 5 mg/kg) administered treatment groups given daily (QD) or every third day (Q3D), when compared to vehicle control (Vehicle) and when IP is compared to IC. Mean Th memory cells were measured using FACS analysis. Mann-Whitney's U-test and Student's t-test were used for statistical analysis on non-Gaussian and Gaussian data respectively. A value of p < 0.05 was considered significant (Graph Pad Software, Inc.).
FIG. 103 is a graph showing the Disease Activity Index (DAI) of naive mice (Group 1), mice administered vehicle only both intraperitoneally (IP) and intracecally (IC) (Group 2), mice administered an anti-TNFa antibody IP and vehicle IC (Group 7), and mice administered an anti-TNFa antibody IC and vehicle IP (Group 8) at Day 28 and Day 42 of the study described in Example 16.
FIG. 104 is a set of graphs showing the colonic tissue concentration of TNFa, IL-17A, IL-4, and IL-22 in mice administered vehicle only both IP and IC (Group 2), mice administered IgG control antibody IP and vehicle IC (Group 3), mice administered IgG
control IC and vehicle IP (Group 4), mice administered anti-TNFa antibody IP
and vehicle IC
(Group 7), and mice administered anti-TNFa antibody IC and vehicle IP (Group 8) at Day 42 of the study described in Example 16.

FIG. 105 is a graph showing the Disease Activity Index (DAI) of naive mice (Group 1), mice administered vehicle only both IP and IC (Group 2), mice administered an anti-IL12 p40 antibody IP and vehicle IC (Group 5), and mice administered an anti-IL12 p40 antibody IC and vehicle IP (Group 6) at Day 28 and Day 42 of the study described in Example 16.
FIG. 106 is a set of graphs showing the colonic tissue concentration of IFNgamma, IL-6, IL-17A, TNFa, IL-22, and IL-lb in naïve mice (Group 1), mice administered vehicle only both IP and IC (Group 2), mice administered anti-IL12 p40 antibody IP and vehicle IC
(Group 5), and mice administered anti-IL12 p40 antibody IC and vehicle IP
(Group 8) at Day 42 of the study described in Example 16.
DETAILED DESCRIPTION
The present disclosure is directed to various methods and formulations for treating diseases of the gastrointestinal tract with a therapeutic agent as disclosed herein. For example, in an embodiment, a method of treating a disease of the gastrointestinal tract in a subject comprises administering to the subject a pharmaceutical formulation comprising a therapeutic agent as disclosed herein wherein the pharmaceutical formulation is released in the subject's gastrointestinal tract proximate to one or more sites of disease. For example, in an embodiment, the pharmaceutical formulation comprises a therapeutically effective amount of a therapeutic agent as disclosed herein.
In some embodiments, the formulation is contained in an ingestible device, and the device releases the formulation at a location proximate to the site of disease. The location of the site of disease may be predetermined. For example, an ingestible device, the location of which within the GI tract can be accurately determined as disclosed herein, may be used to sample one or more locations in the GI tract and to detect one or more analytes, including markers of the disease, in the GI tract of the subject. A pharmaceutical formulation may be then administered via an ingestible device and released at a location proximate to the predetermined site of disease. The release of the formulation may be triggered autonomously, as further described herein.
The following disclosure illustrates aspects of the formulations and methods embodied in the claims.

Formulations, including Pharmaceutical Formulations As used herein, a "formulation" of an immune modulator may refer to either the immune modulator in pure form ¨ such as, for example, the lyophilized immune modulator ¨
or a mixture of the immune modulator with one or more physiologically acceptable carriers, excipients or stabilizers. Thus, therapeutic formulations or medicaments can be prepared by mixing the immune modulator having the desired degree of purity with optional physiologically acceptable carriers, excipients or stabilizers (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the form of lyophilized formulations or aqueous solutions. Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine;
preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride;
benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) antibody; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine;
monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol;
salt- forming counter-ions such as sodium; metal complexes (e.g., Zn-protein complexes);
and/or non-ionic surfactants such as TWEENTm, PLURONICSTM or polyethylene glycol (PEG).
Exemplary pharmaceutically acceptable carriers herein further include insterstitial drug dispersion agents such as soluble neutral-active hyaluronidase glycoproteins (sHASEGP), for example, human soluble PH-20 hyaluronidase glycoproteins, such as rHuPH20 (HYLENEX , Baxter International, Inc.). Certain exemplary sHASEGPs and methods of use, including rHuPH20, are described in US Patent Publication Nos. 2005/0260186 and 2006/0104968. In one aspect, a sHASEGP is combined with one or more additional glycosaminoglycanases such as chondroitinases. Exemplary lyophilized formulations are described in US Patent No. 6,267,958. Aqueous formulations include those described in US Patent No.
6,171,586 .. and W02006/044908, the latter formulations including a histidine-acetate buffer.
A formulation of an immune modulator as disclosed herein, e.g., sustained-release formulations, can further include a mucoadhesive agent, e.g., one or more of polyvinyl pyrolidine, methyl cellulose, sodium carboxyl methyl cellulose, hydroxyl propyl cellulose, carbopol, a polyacrylate, chitosan, a eudragit analogue, a polymer, and a thiomer. Additional examples of mucoadhesive agents that can be included in a formulation with a therapeutic agent as disclosed herein are described in, e.g., Peppas et al., Biomaterials 17(16):1553-1561, 1996; Kharenko et al., Pharmaceutical Chemistry I 43(4):200-208, 2009; Salamat-Miller et al., Adv. Drug Deliv. Reviews 57(11):1666-1691, 2005; Bernkop-Schnurch, Adv.
Drug Deliv.
Rev. 57(11):1569-1582, 2005; and Harding et al., Biotechnol. Genet. Eng. News 16(1):41-86, 1999.
In some embodiments, components of a formulation may include any one of the following components, or any combination thereof: Acacia, Alginate, Alginic Acid, .. Aluminum Acetate, an antiseptic, Benzyl Alcohol, Butyl Paraben, Butylated Hydroxy Toluene, an antioxidant. Citric acid, Calcium carbonate, Candelilla wax, a binder, Croscarmellose sodium, Confectioner sugar, Colloidal silicone dioxide, Cellulose, Carnuba wax, Corn starch, Carboxymethylcellulose calcium, Calcium stearate, Calcium disodium EDTA, Chelation agents, Copolyvidone, Castor oil hydrogenated, Calcium hydrogen phosphate dehydrate, Cetylpyridine chloride, Cysteine HC1, Crosspovidone, Dibasic Calcium Phosphate, Disodium hydrogen phosphate, Dimethicone, Erythrosine Sodium, Ethyl Cellulose, Gelatin, Glyceryl monooleate, Glycerin, Glycine, Glyceryl monostearate, Glyceryl behenate, Hydroxy propyl cellulose, Hydroxyl propyl methyl cellulose, Hypromellose, HPMC Pthalate, Iron oxides or ferric oxide, Iron oxide yellow, Iron oxide red or ferric oxide, .. Lactose (hydrous or anhydrous or monohydrate or spray dried), Magnesium stearate, Microcrystalline cellulose, Mannitol, Methyl celluloseõ Magnesium carbonate, Mineral oil, Methacrylic acid copolymer, Magnesium oxide, Methyl paraben, PEG, Polysorbate 80, Propylene glycol, Polyethylene oxide, Propylene paraben, Polaxamer 407 or 188 or plain, Potassium bicarbonate, Potassium sorbate, Potato starch, Phosphoric acid, Polyoxy140 stearate, Sodium starch glycolate, Starch pregelatinized, Sodium crossmellose, Sodium lauryl sulfate, Starch, Silicon dioxide, Sodium benzoateõ Stearic acid, Sucrose base for medicated confectionery, a granulating agent, Sorbic acid, Sodium carbonate, Saccharin sodium, Sodium alginate, Silica gel, Sorbiton monooleate, Sodium stearyl fumarate, Sodium chloride, Sodium metabisulfite, Sodium citrate dehydrate, Sodium starch, Sodium carboxy methyl cellulose, Succinic acid, Sodium propionate, Titanium dioxide, Talc, Triacetin, Triethyl citrate.
Accordingly, in some embodiments of the method of treating a disease as disclosed herein, the method comprises administering to the subject a pharmaceutical composition that is a formulation as disclosed herein. In some embodiments the formulation is a dosage form, which may be, as an example, a solid form such as, for example, a capsule, a tablet, a sachet, or a lozenge; or which may be, as an example, a liquid form such as, for example, a solution, a suspension, an emulsion, or a syrup.
In some embodiments the formulation is not comprised in an ingestible device.
In some embodiments wherein the formulation is not comprised in an ingestible device, the formulation may be suitable for oral administration. The formulation may be, for example, a solid dosage form or a liquid dosage form as disclosed herein. In some embodiments wherein the formulation is not comprised in an ingestible device, the formulation may be suitable for rectal administration. The formulation may be, for example, a dosage form such as a suppository or an enema. In embodiments where the formulation is not comprised in an ingestible device, the formulation releases the immune modulator at a location in the gastrointestinal tract of the subject that is proximate to an intended site of release in the GI
tract. Such localized release may be achieved, for example, with a formulation comprising an enteric coating. Such localized release may be achieved, an another example, with a formulation comprising a core comprising one or more polymers suitable for controlled release of an active substance. A non-limiting list of such polymers includes:
poly(2-(diethylamino)ethyl methacrylate, 2-(dimethylamino)ethyl methacrylate, poly(ethylene glycol), poly(2-aminoethyl methacrylate), (2-hydroxypropyl)methacrylamide, poly(f3-benzyl-1-aspartate), poly(N-isopropylacrylamide), and cellulose derivatives.
In some embodiments the formulation is comprised in an ingestible device as disclosed herein. In some embodiments wherein the formulation is comprised in an ingestible device, the formulation may be suitable for oral administration.
The formulation may be, for example, a solid dosage form or a liquid dosage form as disclosed herein. In some embodiments the formulation is suitable for introduction and optionally for storage in the device. In some embodiments the formulation is suitable for introduction and optionally for storage in the reservoir comprised in the device. In some embodiments the formulation is suitable for introduction and optionally for storage in the reservoir comprised in the device.
Thus, in some embodiments, provided herein is a reservoir comprising a therapeutically effective amount of an immune modulator, wherein the reservoir is configured to fit into an ingestible device. In some embodiments, the reservoir comprising a therapeutically effective amount of an immune modulator is attachable to an ingestible device. In some embodiments, the reservoir comprising a therapeutically effective amount of an immune modulator is capable of anchoring itself to the subject's tissue. As an example, the reservoir capable of anchoring itself to the subject's tissue comprises silicone. As an example, the reservoir capable of anchoring itself to the subject's tissue comprises polyvinyl chloride.
In some embodiments the formulation is suitable for introduction in the spray catheters disclosed herein.
The formulation/medicament herein may also contain more than one active compound as necessary for the particular indication being treated, for example, those with complementary activities that do not adversely affect each other. For instance, the formulation may further comprise another immune modulator or a chemotherapeutic agent.
Such molecules are suitably present in combination in amounts that are effective for the purpose intended.
The active ingredients may also be entrapped in microcapsule prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsule and poly-(methylmethacylate) microcapsule, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980).
The formulations to be used for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes.
Sustained-release preparations may be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the immune modulator, which matrices are in the form of shaped articles, e.g., films, or microcapsule. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2- hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides (U.S.
Pat. No. 3,773,919), copolymers of L-glutamic acid and y ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON
DEPOTTm (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(-)-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydrogels release proteins for shorter time periods. When encapsulated immune modulators remain in the body for a long time, they may denature or aggregate as a result of exposure to moisture at 37 C, resulting in a loss of biological activity and possible changes in immunogenicity. Rational strategies can be devised for stabilization depending on the mechanism involved. For example, if the aggregation mechanism is discovered to be intermolecular S-S bond formation through thio-disulfide interchange, stabilization may be achieved by modifying sulfhydryl residues, lyophilizing from acidic solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions.
Pharmaceutical formulations may contain one or more immune modulators. The pharmaceutical formulations may be formulated in any manner known in the art.
In some embodiments the formulations include one or more of the following components:
a sterile diluent (e.g., sterile water or saline), a fixed oil, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents, antibacterial or antifungal agents, such as benzyl alcohol or methyl parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like, antioxidants, such as ascorbic acid or sodium bisulfite, chelating agents, such as ethylenediaminetetraacetic acid, buffers, such as acetates, citrates, or phosphates, and isotonic agents, such as sugars (e.g., dextrose), polyalcohols (e.g., mannitol or sorbitol), or salts (e.g., sodium chloride), or any combination thereof. Liposomal suspensions can also be used as pharmaceutically acceptable carriers (see, e.g., U.S. Patent No. 4,522,811, incorporated by reference herein in its entirety). The formulations can be formulated and enclosed in ampules, disposable syringes, or multiple dose vials. Where required, proper fluidity can be maintained by, for example, the use of a coating, such as lecithin, or a surfactant. Controlled release of the immune modulator can be achieved by implants and microencapsulated delivery systems, which can include biodegradable, biocompatible polymers (e.g., ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid; Alza Corporation and Nova Pharmaceutical, Inc.).
In some embodiments, the immune modulator is present in a pharmaceutical formulation within the device.
In some embodiments, the immune modulator is present in solution within the device.
In some embodiments, the immune modulator is present in a suspension in a liquid medium within the device.
In some embodiments, the therapeutic agent as disclosed herein is present as a pure, powder (e.g., lyophilized) form of the therapeutic agent as disclosed herein.

Definitions By "ingestible," it is meant that the device can be swallowed whole.
The terms "antibody" and "immunoglobulin" are used interchangeably in the broadest sense and include monoclonal antibodies (for example, full length or intact monoclonal antibodies), polyclonal antibodies, multivalent antibodies, multi specific antibodies (e.g., bispecific, trispecific etc. antibodies so long as they exhibit the desired biological activity) and may also include certain antibody fragments (as described in greater detail herein). An antibody can be human, humanized and/or affinity matured.
"Antibody fragments" comprise only a portion of an intact antibody, where in certain embodiments, the portion retains at least one, and typically most or all, of the functions normally associated with that portion when present in an intact antibody. In one embodiment, an antibody fragment comprises an antigen binding site of the intact antibody and thus retains the ability to bind antigen. In another embodiment, an antibody fragment, for example one that comprises the Fc region, retains at least one of the biological functions normally associated with the Fc region when present in an intact antibody, such as FcRn binding, antibody half-life modulation, ADCC function and complement binding. In one embodiment, an antibody fragment is a monovalent antibody that has an in vivo half-life substantially similar to an intact antibody. For example, such an antibody fragment may comprise on antigen binding arm linked to an Fc sequence capable of conferring in vivo stability to the fragment.
The term "monoclonal antibody" as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigen. Furthermore, in contrast to polyclonal antibody preparations that typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen.
The monoclonal antibodies herein specifically include "chimeric" antibodies in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Patent No.
4,816,567; and Morrison et al, Proc. Natl. Acad. Sci. USA 81 :6851-6855 (1984)).
"Treatment regimen" refers to a combination of dosage, frequency of administration, or duration of treatment, with or without addition of a second medication.
"Effective treatment regimen" refers to a treatment regimen that will offer beneficial response to a patient receiving the treatment.
"Effective amount" refers to an amount of drug that offers beneficial response to a patient receiving the treatment. For example, an effective amount may be a Human Equivalent Dose (HED).
"Dispensable," with reference to any substance, refers to any substance that may be released from an ingestible device as disclosed herein, or from a component of the device such as a reservoir. For example, a dispensable substance may be a therapeutic agent as disclosed herein, and/or a formulation comprising a therapeutic agent as disclosed herein.
"Patient response" or "patient responsiveness" can be assessed using any endpoint indicating a benefit to the patient, including, without limitation, (1) inhibition, to some extent, of disease progression, including slowing down and complete arrest; (2) reduction in the number of disease episodes and/or symptoms; (3) reduction in lesional size;
(4) inhibition (i.e., reduction, slowing down or complete stopping) of disease cell infiltration into adjacent peripheral organs and/or tissues; (5) inhibition (i.e., reduction, slowing down or complete stopping) of disease spread; (6) decrease of auto-immune response, which may, but does not have to, result in the regression or ablation of the disease lesion; (7) relief, to some extent, of one or more symptoms associated with the disorder; (8) increase in the length of disease-free presentation following treatment; and/or (9) decreased mortality at a given point of time following treatment. The term "responsiveness" refers to a measurable response, including complete response (CR) and partial response (PR).
As used herein, "complete response" or "CR" means the disappearance of all signs of inflammation or remission in response to treatment. This does not necessarily mean the disease has been cured.
"Partial response" or "PR" refers to a decrease of at least 50% in the severity of inflammation, in response to treatment.
A "beneficial response" of a patient to treatment with a therapeutic agent and similar wording refers to the clinical or therapeutic benefit imparted to a patient at risk for or suffering from a inflammatory disease or condition that arises in a tissue originating from the endoderm. Such benefit includes cellular or biological responses, a complete response, a partial response, a stable disease (without progression or relapse), or a response with a later relapse of the patient from or as a result of the treatment with the agent.
As used herein, "non-response" or "lack of response" or similar wording means an absence of a complete response, a partial response, or a beneficial response to treatment with a therapeutic agent.
"A patient maintains responsiveness to a treatment" when the patient' s responsiveness does not decrease with time during the course of a treatment.
A "symptom" of a disease or disorder (e.g., an inflammatory disease or condition that arises in tissue originating from the endoderm) is any morbid phenomenon or departure from the normal in structure, function, or sensation, experienced by a subject and indicative of disease.
"Mucosa-associated lymphoid tissue" or "MALT" refers to a diffuse system of small concentrations of lymphoid tissue found in various submucosal membrane sites of the body, such as the gastrointestinal tract, oral passage, nasopharyngeal tract, thyroid, breast, lung, salivary glands, eye, and skin.
"Gut-associated lymphoid tissue" or "GALT" refers to a part of the broader MALT
and includes, e.g., Peyer's patches, mesenertic lymph nocdes, and isolated lymphoid follicles/intestinal lymphoid aggregates.
"Peyer's patches" refers to aggregated lymphoid modules organized into follicles and are important part of GALT. Peyer's patches are mainly present in the distal jejunum and the ileum.
"Mesenteric lymph nodes" refers to part of the paraaortic lymph node system that is a group of lymph nodes that lie between the layers of the mesentery and drain the gut tissues and deliver lymph to the thoracic duct. Mesenteric lymph nodes include the "superior mesenteric lymph nodes" which receive afferents from the jejunum, ileum, cecum, and the ascending and parts of the transverse colon. Mesenteric lymph nodes also include "inferior mesenteric lymph nodes" which are lymph nodes present throughout the hindgut.
The hindgut, e.g., includes the distal third of the transverse colon and the splenic flexure, the descending colon, sigmoid colon, and the rectum. The lymph nodes drain into the superior mesenteric lymph nodes and ultimately to the preaortic lymph nodes.
"Paraaortic lymph nodes" refers to a group of mesenteric lymph nodes that lie in front of the lumbar vertebrae near the aorta. The paraaortic lymph nodes receive drainage from the gastrointestinal tract and the abdominal organs. Paraaortic lymph nodes include, e.g., retroaortic lymph nodes, lateral aortic lymph nodes, preaortic lymph nodes (e.g., Celiac, gastic, hepatic, and splenic lymph nodes), superior mesenteric lymph nodes (e.g., mesenteric, ileocolic, and mesocolic lymph nodes), and inferior mesenteric lymph nodes (e.g., pararectal lymph nodes).
As used herein, "accuracy," when disclosed in connection with a specified location of a device within the GI tract of a subject, refers to the degree to which the location determined by the device conforms to the correct location, wherein the correct location is based on a generally accepted standard. The location within the GI tract of the subject determined by the device can be based on data, for example, light reflectance data, collected by the ingestible device. In some embodiments, the correct location can be based on external imaging devices, such as computer-aided tomography (CT), interpreted, for example, by a qualified clinician or physician. Therefore, percent accuracy ("% accuracy") can refer to the percentage agreement between the location of the device in the GI tract as determined by the device, and the correct location, for example, as determined by CT, e.g., expressed as [(number of devices in which location determined by the device agrees with location as determined by CT /
total devices administered to the subject or subjects) x 100%], or, where only one device is administered per subject, [(number of subjects in which location determined by the device agrees with location as determined by CT / total number of subjects) x 100%]. The latter formula for determining % accuracy was used in Example 14. In some embodiments, the accuracy with which the device determines a location refers to the accuracy with which the device determines that it is at a location pre-selected for drug release.
As used herein, an "autonomous device" refers to a device comprising one or more processors configured to independently control certain mechanisms or operations of the device while in the GI tract of a subject. Preferably, an autonomous device of the invention has no external electrical or wireless connections that control device mechanisms or operations, although connections such as wireless connections may be present to enable alternative device functions, such as transmitting data collected by the device to an external (ex vivo) system or receiver. The independently controlled mechanisms or operations of the autonomous device include, for example, triggering the release of a drug (or the formulation comprising the drug), triggering collection of one or more samples, and/or triggering the analysis of one or more samples; and/or determining the location of the device within the GI
tract of the subject. Such a mechanism is referred to herein as an "autonomous mechanism,"

for example, an "autonomous triggering mechanism" or an "autonomous localization mechanism," respectively. Actively implementing such an autonomous triggering or localization mechanism is referred to as "autonomous triggering" or "autonomous localizing," respectively. An "autonomous localization mechanism" is synonymous with a "self-localization mechanism.
As used herein, a "housing" is a portion of an ingestible device that defines the boundary between the interior of the device and the environment exterior to the device.
As used herein, a "self-localizing device" refers to a device comprising a mechanism or system that can be implemented autonomously to determine the location of the ingestible device in vivo, e.g., within the GI tract of a subject. Such a mechanism is referred to as a "self-localization mechanism." A "self-localization mechanism" is synonymous with an "autonomous localization mechanism."A self-localizing device does not require ex vivo visualization devices or systems, for example, using scintigraphy or computer-aided tomography (CT), to localize in the GI tract.
As used herein, "localizing the device" refers to determining a location of the device.
As used herein, "sensor" refers to a mechanism or portion of a mechanism configured to collect information regarding the surroundings of the ingestible device.
Examples of "sensors" include environmental sensors and light sensors. Examples of environmental sensors include pH sensors and sensors capable to identifying muscle contractions and/or peristalsis.
As used herein, "time following transition" refers to elapsed time after passage of the device from one portion, section or subsection of the GI tract into an adjacent portion, section or subsection of the GI tract.
As used herein, "proximate" as disclosed in connection with release of a drug from a device to one or more disease sites, refers to a location that is sufficiently spatially close to the one or more disease sites such that releasing the drug at the location treats the disease.
For example, when the drug is released proximate to the one or more disease sites, the drug may be released 150 cm or less, such as 125 cm or less, such as 100 cm or less, such as 50 cm or less, such as 40 cm or less, such as 30 cm or less, such as 20 cm or less, such as 10 cm or less, such as 5 cm or less, such as 2 cm or less, from the one or more sites of disease. The proximate location for drug release may be in the same section or subsection of the gastrointestinal tract as the one or more disease sites. In the alternative, the proximate location for drug release may be in a different section or subsection of the GI tract than the one or more disease sites; for example, the drug release may be proximal to the one or more disease sites. In a non-limiting example, the drug may be released in the cecum to treat a site of disease tissue in the ascending colon (i.e., distal to the cecum). In another non-limiting example, the drug may be released in the cecum to treat a site of disease tissue in one or more of the ascending colon, transverse colon, descending colon, or rectum. Thus, where the present application refers to release of a drug proximate to a site of disease, this may in some embodiments refer to release in a section or subsection of the GI tract which has been determined to contain a site of disease. The section may be selected from esophagus, stomach, duodenum, jejunum, ileum, cecum, ascending colon, transverse colon, descending colon, and rectum. The subsection may be selected from proximal duodenum, proximal jejunum, proximal ileum, proximal cecum, proximal ascending colon, proximal transverse colon, proximal descending colon, distal duodenum, distal jejunum, distal ileum, distal cecum, distal ascending colon, distal transverse colon, distal descending colon.
As used herein, the "total induction dose" is the sum of induction doses over a given time period.
As used herein, "proximal", when used in connection with an anatomical structure, refers to a portion, section, or subsection that precedes, or is upstream of, an adjacent portion, section, or subsection of the anatomical structure. In some embodiments, proximal refers to a portion, section, or subsection that immediately precedes, or is immediately upstream of, an immediately adjacent portion, section, or subsection of the anatomical structure.
As used herein, "distal", when used in connection with an anatomical structure, refers to a portion, section, or subsection that follows, or is downstream of, an adjacent portion, section, or subsection of the anatomical structure. In some embodiments, distal refers to a portion, section, or subsection that immediately follows, or is immediately downstream of, an immediately adjacent portion, section, or subsection of the anatomical structure.
As used herein, a reference to a drug's international nonproprietary name (INN) is to be interpreted as including generic, bioequivalent and biosimilar versions of that drug, including but not limited to any drug that has received abbreviated regulatory approval by reference to an earlier regulatory approval of that drug.

Inflammatory Conditions or Diseases that Arise from a Tissue Originating from the Endoderm The presently claimed devices can, e.g., provide for a higher concentration of a4137 expressing cells in the periphery (e.g., blood) when an immune modulator is delivered topically to one or more parts of the GI tract distal to the stomach (e.g., the small or large intestine) as compared to when the same dose of the immune modulator is systemically administered. The presently claimed devices can, e.g., result in trafficked cells being forced out of the local gastrointestinal tissue (including the mucosa) and lymph system, and back into systemic circulation of a subject.
Accordingly, also provided herein are methods of treating a disease or condition that arises in a tissue originating from the endoderm. The endoderm forms the gastrointestinal tract, respiratory tract, endocrine glands, and organs, the auditory system and urinary system.
Thus, the present invention includes compositions and devices for treating diseases and conditions found in the following tissues that originate from the endoderm (e.g., the stomach, the colon, the liver, the pancreas, the urinary bladder, the epithelial parts of the trachea, the lungs, the pharynx, the thyroid, the parathyroid, the intestines, and the gallbladder). Also provided herein are methods of treating a disease or a condition that arises in a tissue originating from the endoderm (e.g., any of the exemplary diseases or conditions that arise in a tissue originating from the endoderm described herein) that include intrathecally releasing one or more immune modulators in the small or large intestine using any of the devices or compositions described herein.
Non-limiting examples of a disease or condition that arises in a tissue originating from the endoderm includes gastritis, Celiac disease, hepatitis, alcoholic lever disease, fatty liver disease (hepatic steatosis), non-alcoholic fatty liver disease (NASH), cirrhosis, primary schlerosing cholangitis, pancreatitis, insterstitial cystitits, asthma, chronic obstructic pulmonary disease, pulmonary fibrosis, pharyngitis, thyroiditis, hyperthyroidism, parathyroiditis, nephritis, Hashimoto's disease, Addison's disease, Graves' disease, Sjogren syndrome, type 1 diabetes, pelvic inflammatory disease, auditory canal inflammation, tinnitus, vestibular neuritis, otitis media, auditory canal inflammation, tracheitis, cholestatic liver disease, primary biliary schlerosis, liver parenchyma, an inherited metabolic disorder of the liver, Byler syndrome, cerebrotendinous, xanthomatosis, Zellweger's syndrome, neonatal hepatitis, cystic fibrosis, ALGS (Alagilles syndrome), PFIC (progressive familial intrahepatic cholestasis), autoimmune hepatitis, primary biliary cirrhosis (PBC), liver fibrosis, NAFLD, portal hypertension, general cholestasis, such as in jaundice due to drugs or during pregnancy, intra- and extrahepatic cholestasis, such as hereditary forms of cholestasis, such as PFIC1, gall stones and choledocholithiasis, malignancy causing obstruction of the biliary tree, symptoms (scratching, pruritus) due to cholestasis/jaundice, chronic autoimmune liver disease leading to progressive cholestasis, and pruritus of cholestatic liver disease, duodenal ulcers, enteritis (radition-, chemotherapy-, or infection-induced enteritis), diverticulitis, pouchitis, cholecystitis, and cholangitis. Additional examples of diseases and conditions that arise in a tissue originating from the endoderm are known in the are known in the art.
As used herein, the term "immune modulator" means a therapeutic agent that .. decreases the activation of an immune cell (e.g., a T cell, e.g., memory T
cell), decreases the secretion or expression of a pro-inflammatory cytokine, decreases the recruitment or migration of T-lymphocytes (e.g., memory T lymphocytes), and/or increases the secretion or expression of an anti-inflammatory cytokine. Non-limiting examples of immune modulators are anti-inflammatory agents. Non-limiting examples of anti-inflammatory agents include IL-12/IL-23 inhibitors, TNFa inhibitors, IL-6 receptor inhibitors, immune modulatory agents (e.g., CD40/CD4OL inhibitors), IL-1 inhibitors, IL-13 inhibitors, IL-10 receptor agonists, chemokine/chemokine receptor inhibitors, and integrin inhibitors. Non-limiting examples of integrin inhibitors include 137 integrin inhibitors, such as a4137 integrin inhibitors. In some embodiments of any of the methods described herein, the immune modulator is a .. inhibitor.
As used herein, the term "immune modulator" means a therapeutic agent that decreases the activation of an immune cell, decreases the secretion or expression of a pro-inflammatory cytokine, decreases the recruitment or migration of T-lymphocytes (e.g., memory T lymphocytes), and/or increases the secretion or expression of an anti-inflammatory cytokine. Non-limiting examples of immune modulators are anti-inflammatory agents. Non-limiting examples of anti-inflammatory agents include IL-12/IL-23 inhibitors, TNFa inhibitors, IL-6 receptor inhibitors, immune modulatory agents (e.g., inhibitors), IL-1 inhibitors, IL-13 inhibitors, IL-10 receptor agonists, chemokine/chemokine receptor inhibitors, and integrin inhibitors. In some embodiments of any of the methods .. described herein, the immune modulator is a PDE4 inhibitor. Additional examples of immune modulators useful for the treatment of a liver disease or disorder are described below.

Non-limiting exemplary examples of immune modulators are described below.
Additional examples of immune modulators are known in the art.
IL-12/ IL-23 Inhibitors The term "IL-12/IL-23 inhibitors" refers to an agent which decreases IL-12 or expression and/or the ability of IL-12 to bind to an IL-12 receptor or the ability of IL-23 to bind to an IL-23 receptor. IL-12 is a heterodimeric cytokine that includes both IL-12A (p35) and IL-12B (p40) polypeptides. IL-23 is a heterodimeric cytokine that includes both IL-23 (p19) and IL-12B (p40) polypeptides. The receptor for IL-12 is a heterodimeric receptor includes IL-12R 131 and IL-12R 132. The receptor for IL-23 receptor is a heterodimeric receptor that includes both IL-12R 131 and IL-23R.
In some embodiments, the IL-12/IL-23 inhibitor can decrease the binding of IL-12 to the receptor for IL-12. In some embodiments, the IL-12/IL-23 inhibitor can decrease the binding of IL-23 to the receptor for IL-23. In some embodiments, the IL-12/IL-23 inhibitor decreases the expression of IL-12 or IL-23. In some embodiments, the IL-12/IL-23 inhibitor decreases the expression of a receptor for IL-12. In some embodiments, the IL-inhibitor decreases the expression of a receptor for IL-23.
In some embodiments, the IL-12/IL-23 inhibitory agent targets IL-12B (p40) subunit.
In some embodiments, the IL-12/IL-23 inhibitory agent targets IL-12A (p35). In some embodiments, the IL-12/IL-23 inhibitory agent targets IL-23 (p19). In some embodiments, the IL-12/IL-23 inhibitory agent targets the receptor for IL-12 (one or both of IL-12R 131 or IL-12R (32). In some embodiments, the IL-12/IL-23 inhibitory agent targets the receptor for IL-23 (one or both of IL-12R 131 and IL-23R).
In some embodiments, an IL-12/IL-23 inhibitor can be an inhibitory nucleic acid. In some embodiments, the inhibitory nucleic acid can be an antisense nucleic acid, a ribozyme, and a small interfering RNA (siRNA). Examples of aspects of these different oligonucleotides are described below. Any of the examples of inhibitory nucleic acids that can decrease expression of IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R
131, IL-12R 132, or IL-23R mRNA in a mammalian cell can be synthesized in vitro.
Inhibitory nucleic acids that can decrease the expression of IL-12A (p35), IL-(p40), IL-23 (p19), IL-12R 131, IL-12R 132, or IL-23R mRNA expression in a mammalian cell include antisense nucleic acid molecules, i.e., nucleic acid molecules whose nucleotide sequence is complementary to all or part of an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R mRNA (e.g., complementary to all or a part of any one of SEQ
ID NOs: 1-12).
Human IL-12A (p35) mRNA (SEQ ID NO: 1) 1 tttcgctttc attttgggcc gagctggagg cggcggggcc gtcccggaac ggctgcggcc 61 gggcaccccg ggagttaatc cgaaagcgcc gcaagccccg cgggccggcc gcaccgcacg 121 tgtcaccgag aagctgatgt agagagagac acagaaggag acagaaagca agagaccaga 181 gtcccgggaa agtcctgccg cgcctcggga caattataaa aatgtggccc cctgggtcag 241 cctcccagcc accgccctca cctgccgcgg ccacaggtct gcatccagcg gctcgccctg 301 tgtccctgca gtgccggctc agcatgtgtc cagcgcgcag cctcctcctt gtggctaccc 361 tggtcctcct ggaccacctc agtttggcca gaaacctccc cgtggccact ccagacccag 421 gaatgttccc atgccttcac cactcccaaa acctgctgag ggccgtcagc aacatgctcc 481 agaaggccag acaaactcta gaattttacc cttgcacttc tgaagagatt gatcatgaag 541 atatcacaaa agataaaacc agcacagtgg aggcctgttt accattggaa ttaaccaaga 601 atgagagttg cctaaattcc agagagacct ctttcataac taatgggagt tgcctggcct 661 ccagaaagac ctcttttatg atggccctgt gccttagtag tatttatgaa gacttgaaga 721 tgtaccaggt ggagttcaag accatgaatg caaagcttct gatggatcct aagaggcaga 781 tctttctaga tcaaaacatg ctggcagtta ttgatgagct gatgcaggcc ctgaatttca 841 acagtgagac tgtgccacaa aaatcctccc ttgaagaacc ggatttttat aaaactaaaa 901 tcaagctctg catacttctt catgctttca gaattcgggc agtgactatt gatagagtga 961 tgagctatct gaatgcttcc taaaaagcga ggtccctcca aaccgttgtc atttttataa 1021 aactttgaaa tgaggaaact ttgataggat gtggattaag aactagggag ggggaaagaa 1081 ggatgggact attacatcca catgatacct ctgatcaagt atttttgaca tttactgtgg 1141 ataaattgtt tttaagtttt catgaatgaa ttgctaagaa gggaaaatat ccatcctgaa 1201 ggtgtttttc attcacttta atagaagggc aaatatttat aagctatttc tgtaccaaag 1261 tgtttgtgga aacaaacatg taagcataac ttattttaaa atatttattt atataacttg 1321 gtaatcatga aagcatctga gctaacttat atttatttat gttatattta ttaaattatt 1381 tatcaagtgt atttgaaaaa tatttttaag tgttctaaaa ataaaagtat tgaattaaag 1441 tgaaaaaaaa Human IL-12B (p40) mRNA (SEQ ID NO: 2) 1 ctgtttcagg gccattggac tctccgtcct gcccagagca agatgtgtca ccagcagttg 61 gtcatctctt ggttttccct ggtttttctg gcatctcccc tcgtggccat atgggaactg 121 aagaaagatg tttatgtcgt agaattggat tggtatccgg atgcccctgg agaaatggtg 181 gtcctcacct gtgacacccc tgaagaagat ggtatcacct ggaccttgga ccagagcagt 241 gaggtcttag gctctggcaa aaccctgacc atccaagtca aagagtttgg agatgctggc 301 cagtacacct gtcacaaagg aggcgaggtt ctaagccatt cgctcctgct gcttcacaaa 361 aaggaagatg gaatttggtc cactgatatt ttaaaggacc agaaagaacc caaaaataag 421 acctttctaa gatgcgaggc caagaattat tctggacgtt tcacctgctg gtggctgacg 481 acaatcagta ctgatttgac attcagtgtc aaaagcagca gaggctcttc tgacccccaa 541 ggggtgacgt gcggagctgc tacactctct gcagagagag tcagagggga caacaaggag 601 tatgagtact cagtggagtg ccaggaggac agtgcctgcc cagctgctga ggagagtctg 661 cccattgagg tcatggtgga tgccgttcac aagctcaagt atgaaaacta caccagcagc 721 ttcttcatca gggacatcat caaacctgac ccacccaaga acttgcagct gaagccatta 781 aagaattctc ggcaggtgga ggtcagctgg gagtaccctg acacctggag tactccacat 841 tcctacttct ccctgacatt ctgcgttcag gtccagggca agagcaagag agaaaagaaa 901 gatagagtct tcacggacaa gacctcagcc acggtcatct gccgcaaaaa tgccagcatt 961 agcgtgcggg cccaggaccg ctactatagc tcatcttgga gcgaatgggc atctgtgccc 1021 tgcagttagg ttctgatcca ggatgaaaat ttggaggaaa agtggaagat attaagcaaa 1081 atgtttaaag acacaacgga atagacccaa aaagataatt tctatctgat ttgctttaaa 1141 acgttttttt aggatcacaa tgatatcttt gctgtatttg tatagttaga tgctaaatgc 1201 tcattgaaac aatcagctaa tttatgtata gattttccag ctctcaagtt gccatgggcc 1261 ttcatgctat ttaaatattt aagtaattta tgtatttatt agtatattac tgttatttaa 1321 cgtttgtctg ccaggatgta tggaatgttt catactctta tgacctgatc catcaggatc 1381 agtccctatt atgcaaaatg tgaatttaat tttatttgta ctgacaactt ttcaagcaag 1441 gctgcaagta catcagtttt atgacaatca ggaagaatgc agtgttctga taccagtgcc 1501 atcatacact tgtgatggat gggaacgcaa gagatactta catggaaacc tgacaatgca 1561 aacctgttga gaagatccag gagaacaaga tgctagttcc catgtctgtg aagacttcct 1621 ggagatggtg ttgataaagc aatttagggc cacttacact tctaagcaag tttaatcttt 1681 ggatgcctga attttaaaag ggctagaaaa aaatgattga ccagcctggg aaacataaca 1741 agaccccgtc tctacaaaaa aaatttaaaa ttagccaggc gtggtggctc atgcttgtgg 1801 tcccagctgt tcaggaggat gaggcaggag gatctcttga gcccaggagg tcaaggctat 1861 ggtgagccgt gattgtgcca ctgcatacca gcctaggtga cagaatgaga ccctgtctca 1921 aaaaaaaaaa tgattgaaat taaaattcag ctttagcttc catggcagtc ctcaccccca 1981 cctctctaaa agacacagga ggatgacaca gaaacaccgt aagtgtctgg aaggcaaaaa 2041 gatcttaaga ttcaagagag aggacaagta gttatggcta aggacatgaa attgtcagaa 2101 tggcaggtgg cttcttaaca gccctgtgag aagcagacag atgcaaagaa aatctggaat 2161 ccctttctca ttagcatgaa tgaacctgat acacaattat gaccagaaaa tatggctcca 2221 tgaaggtgct acttttaagt aatgtatgtg cgctctgtaa agtgattaca tttgtttcct 2281 gtttgtttat ttatttattt atttttgcat tctgaggctg aactaataaa aactcttctt 2341 tgtaatc Human IL-23 (p19) mRNA (SEQ ID NO: 3) 1 aaaacaacag gaagcagctt acaaactcgg tgaacaactg agggaaccaa accagagacg 61 cgctgaacag agagaatcag gctcaaagca agtggaagtg ggcagagatt ccaccaggac 121 tggtgcaagg cgcagagcca gccagatttg agaagaaggc aaaaagatgc tggggagcag 181 agctgtaatg ctgctgttgc tgctgccctg gacagctcag ggcagagctg tgcctggggg 241 cagcagccct gcctggactc agtgccagca gctttcacag aagctctgca cactggcctg 301 gagtgcacat ccactagtgg gacacatgga tctaagagaa gagggagatg aagagactac 361 aaatgatgtt ccccatatcc agtgtggaga tggctgtgac ccccaaggac tcagggacaa 421 cagtcagttc tgcttgcaaa ggatccacca gggtctgatt ttttatgaga agctgctagg 481 atcggatatt ttcacagggg agccttctct gctccctgat agccctgtgg gccagcttca 541 tgcctcccta ctgggcctca gccaactcct gcagcctgag ggtcaccact gggagactca 601 gcagattcca agcctcagtc ccagccagcc atggcagcgt ctccttctcc gcttcaaaat 661 ccttcgcagc ctccaggcct ttgtggctgt agccgcccgg gtctttgccc atggagcagc 721 aaccctgagt ccctaaaggc agcagctcaa ggatggcact cagatctcca tggcccagca 781 aggccaagat aaatctacca ccccaggcac ctgtgagcca acaggttaat tagtccatta 841 attttagtgg gacctgcata tgttgaaaat taccaatact gactgacatg tgatgctgac 901 ctatgataag gttgagtatt tattagatgg gaagggaaat ttggggatta tttatcctcc 961 tggggacagt ttggggagga ttatttattg tatttatatt gaattatgta cttttttcaa 1021 taaagtctta tttttgtggc taaaaaaaa .. Human IL-12R 131 mRNA Variant 1 (SEQ ID NO: 4) 1 ctctttcact ttgacttgcc ttagggatgg gctgtgacac tttacttttt ttcttttttc 61 ttttttttca gtcttttctc cttgctcagc ttcaatgtgt tccggagtgg ggacggggtg 121 gctgaacctc gcaggtggca gagaggctcc cctggggctg tggggctcta cgtggatccg 181 atggagccgc tggtgacctg ggtggtcccc ctcctcttcc tcttcctgct gtccaggcag 241 ggcgctgcct gcagaaccag tgagtgctgt tttcaggacc cgccatatcc ggatgcagac 301 tcaggctcgg cctcgggccc tagggacctg agatgctatc ggatatccag tgatcgttac 361 gagtgctcct ggcagtatga gggtcccaca gctggggtca gccacttcct gcggtgttgc 421 cttagctccg ggcgctgctg ctacttcgcc gccggctcag ccaccaggct gcagttctcc 481 gaccaggctg gggtgtctgt gctgtacact gtcacactct gggtggaatc ctgggccagg 541 aaccagacag agaagtctcc tgaggtgacc ctgcagctct acaactcagt taaatatgag 601 cctcctctgg gagacatcaa ggtgtccaag ttggccgggc agctgcgtat ggagtgggag 661 accccggata accaggttgg tgctgaggtg cagttccggc accggacacc cagcagccca 721 tggaagttgg gcgactgcgg acctcaggat gatgatactg agtcctgcct ctgccccctg 781 gagatgaatg tggcccagga attccagctc cgacgacggc agctggggag ccaaggaagt 841 tcctggagca agtggagcag ccccgtgtgc gttccccctg aaaacccccc acagcctcag 901 gtgagattct cggtggagca gctgggccag gatgggagga ggcggctgac cctgaaagag 961 cagccaaccc agctggagct tccagaaggc tgtcaagggc tggcgcctgg cacggaggtc 1021 acttaccgac tacagctcca catgctgtcc tgcccgtgta aggccaaggc caccaggacc 1081 ctgcacctgg ggaagatgcc ctatctctcg ggtgctgcct acaacgtggc tgtcatctcc 1141 tcgaaccaat ttggtcctgg cctgaaccag acgtggcaca ttcctgccga cacccacaca 1201 gaaccagtgg ctctgaatat cagcgtcgga accaacggga ccaccatgta ttggccagcc 1261 cgggctcaga gcatgacgta ttgcattgaa tggcagcctg tgggccagga cgggggcctt 1321 gccacctgca gcctgactgc gccgcaagac ccggatccgg ctggaatggc aacctacagc 1381 tggagtcgag agtctggggc aatggggcag gaaaagtgtt actacattac catctttgcc 1441 tctgcgcacc ccgagaagct caccttgtgg tctacggtcc tgtccaccta ccactttggg 1501 ggcaatgcct cagcagctgg gacaccgcac cacgtctcgg tgaagaatca tagcttggac 1561 tctgtgtctg tggactgggc accatccctg ctgagcacct gtcccggcgt cctaaaggag 1621 tatgttgtcc gctgccgaga tgaagacagc aaacaggtgt cagagcatcc cgtgcagccc 1681 acagagaccc aagttaccct cagtggcctg cgggctggtg tagcctacac ggtgcaggtg 1741 cgagcagaca cagcgtggct gaggggtgtc tggagccagc cccagcgctt cagcatcgaa 1801 gtgcaggttt ctgattggct catcttcttc gcctccctgg ggagcttcct gagcatcctt 1861 ctcgtgggcg tccttggcta ccttggcctg aacagggccg cacggcacct gtgcccgccg 1921 ctgcccacac cctgtgccag ctccgccatt gagttccctg gagggaagga gacttggcag 1981 tggatcaacc cagtggactt ccaggaagag gcatccctgc aggaggccct ggtggtagag 2041 atgtcctggg acaaaggcga gaggactgag cctctcgaga agacagagct acctgagggt 2101 gcccctgagc tggccctgga tacagagttg tccttggagg atggagacag gtgcaaggcc 2161 aagatgtgat cgttgaggct cagagagggt gagtgactcg cccgaggcta cgtagcacac 2221 acaggagtca catttggacc caaataaccc agagctcctc caggctccag tgcacctgcc 2281 tcctctctgc cccgtgcctg ttgccaccca tcctgcgggg gaaccctaga tgctgccatg 2341 aaatggaagc tgctgcaccc tgctgggcct ggcatccgtg gggcaggagc agaccctgcc 2401 atttacctgt tctggcgtag aatggactgg gaatgggggc aaggggggct cagatggatc 2461 cctggaccct gggctgggca tccaccccca ggagcactgg atggggagtc tggactcaag 2521 ggctccctgc agcattgcgg ggtcttgtag cttggaggat ccaggcatat agggaagggg 2581 gctgtaaact ttgtgggaaa aatgacggtc ctcccatccc accccccacc ccaccctcac 2641 ccccctataa aatgggggtg gtgataatga ccttacacag ctgttcaaaa tcatcgtaaa 2701 tgagcctcct cttgggtatt tttttcctgt ttgaagcttg aatgtcctgc tcaaaatctc 2761 aaaacacgag ccttggaatt caaaaaaaaa aaaaaaaaaa Human IL-12R 131 mRNA Variant 2 (SEQ ID NO: 5) 1 ctctttcact ttgacttgcc ttagggatgg gctgtgacac tttacttttt ttcttttttc 61 ttttttttca gtcttttctc cttgctcagc ttcaatgtgt tccggagtgg ggacggggtg 121 gctgaacctc gcaggtggca gagaggctcc cctggggctg tggggctcta cgtggatccg 181 atggagccgc tggtgacctg ggtggtcccc ctcctcttcc tcttcctgct gtccaggcag 241 ggcgctgcct gcagaaccag tgagtgctgt tttcaggacc cgccatatcc ggatgcagac 301 tcaggctcgg cctcgggccc tagggacctg agatgctatc ggatatccag tgatcgttac 361 gagtgctcct ggcagtatga gggtcccaca gctggggtca gccacttcct gcggtgttgc 421 cttagctccg ggcgctgctg ctacttcgcc gccggctcag ccaccaggct gcagttctcc 481 gaccaggctg gggtgtctgt gctgtacact gtcacactct gggtggaatc ctgggccagg 541 aaccagacag agaagtctcc tgaggtgacc ctgcagctct acaactcagt taaatatgag 601 cctcctctgg gagacatcaa ggtgtccaag ttggccgggc agctgcgtat ggagtgggag 661 accccggata accaggttgg tgctgaggtg cagttccggc accggacacc cagcagccca 721 tggaagttgg gcgactgcgg acctcaggat gatgatactg agtcctgcct ctgccccctg 781 gagatgaatg tggcccagga attccagctc cgacgacggc agctggggag ccaaggaagt 841 tcctggagca agtggagcag ccccgtgtgc gttccccctg aaaacccccc acagcctcag 901 gtgagattct cggtggagca gctgggccag gatgggagga ggcggctgac cctgaaagag 961 cagccaaccc agctggagct tccagaaggc tgtcaagggc tggcgcctgg cacggaggtc 1021 acttaccgac tacagctcca catgctgtcc tgcccgtgta aggccaaggc caccaggacc 1081 ctgcacctgg ggaagatgcc ctatctctcg ggtgctgcct acaacgtggc tgtcatctcc 1141 tcgaaccaat ttggtcctgg cctgaaccag acgtggcaca ttcctgccga cacccacaca 1201 gatggcatga tctcagctca ctgcaacctc cgccttccag attcaagaga ttctcctgct 1261 tcagcctccc gagtagctgg gattacaggc atctgccacc atacccggct aattttgtat 1321 ttttagtaga gacggggttt caccacgttg gccaggctgg tctcgaactc ctgacctcaa 1381 gtgatccacc tgccttggcc tcccaaagtg ttgggattat aggcgtgagc caccatgccc 1441 agcctaattt ttgtattttt agtagagatg gagtttcacc atgttgccca ggctggtctc 1501 aaactcctgc cctcaggtga tccacccacc tcagcctctc aaagtgctgg gattacaggt 1561 gtgagccact gtggccgacc tactattttt attatttttg agctaggttc tcagtctgtt 1621 ggcagactgg agtgcaatca tggctcactg cagccttgaa ctcccagact caagtgatcc 1681 ttccacctca gcctctggag tagctgggac tacagacatg caccaccaca cctggttaat 1741 tttttatttt tattttttgt agagacaggt gtctctctac gttgcccagg ctggtctcga 1801 actcctgggc tcaagtgatc cacccatctc cacctcccaa agtgctagga ttacaggcgt 1861 gagccaccgt acccagcctg gtcccatatc atagtgaaat ggtgcctgta aagctctcag 1921 cattggcttg gcacatgcag ttggtactca ataaacggct gttgctatcc ccaaaaaaaa 1981 aaaaaaaaaa aaaaaaa Human IL-12R 131 mRNA Variant 3 (SEQ ID NO: 6) 1 ctctttcact ttgacttgcc ttagggatgg gctgtgacac tttacttttt ttcttttttc 61 ttttttttca gtcttttctc cttgctcagc ttcaatgtgt tccggagtgg ggacggggtg 121 gctgaacctc gcaggtggca gagaggctcc cctggggctg tggggctcta cgtggatccg 181 atggagccgc tggtgacctg ggtggtcccc ctcctcttcc tcttcctgct gtccaggcag 241 ggcgctgcct gcagaaccag tgagtgctgt tttcaggacc cgccatatcc ggatgcagac 301 tcaggctcgg cctcgggccc tagggacctg agatgctatc ggatatccag tgatcgttac 361 gagtgctcct ggcagtatga gggtcccaca gctggggtca gccacttcct gcggtgttgc 421 cttagctccg ggcgctgctg ctacttcgcc gccggctcag ccaccaggct gcagttctcc 481 gaccaggctg gggtgtctgt gctgtacact gtcacactct gggtggaatc ctgggccagg 541 aaccagacag agaagtctcc tgaggtgacc ctgcagctct acaactcagt taaatatgag 601 cctcctctgg gagacatcaa ggtgtccaag ttggccgggc agctgcgtat ggagtgggag 661 accccggata accaggttgg tgctgaggtg cagttccggc accggacacc cagcagccca 721 tggaagttgg gcgactgcgg acctcaggat gatgatactg agtcctgcct ctgccccctg 781 gagatgaatg tggcccagga attccagctc cgacgacggc agctggggag ccaaggaagt 841 tcctggagca agtggagcag ccccgtgtgc gttccccctg aaaacccccc acagcctcag 901 gtgagattct cggtggagca gctgggccag gatgggagga ggcggctgac cctgaaagag 961 cagccaaccc agctggagct tccagaaggc tgtcaagggc tggcgcctgg cacggaggtc 1021 acttaccgac tacagctcca catgctgtcc tgcccgtgta aggccaaggc caccaggacc 1081 ctgcacctgg ggaagatgcc ctatctctcg ggtgctgcct acaacgtggc tgtcatctcc 1141 tcgaaccaat ttggtcctgg cctgaaccag acgtggcaca ttcctgccga cacccacaca 1201 gaaccagtgg ctctgaatat cagcgtcgga accaacggga ccaccatgta ttggccagcc 1261 cgggctcaga gcatgacgta ttgcattgaa tggcagcctg tgggccagga cgggggcctt 1321 gccacctgca gcctgactgc gccgcaagac ccggatccgg ctggaatggc aacctacagc 1381 tggagtcgag agtctggggc aatggggcag gaaaagtgtt actacattac catctttgcc 1441 tctgcgcacc ccgagaagct caccttgtgg tctacggtcc tgtccaccta ccactttggg 1501 ggcaatgcct cagcagctgg gacaccgcac cacgtctcgg tgaagaatca tagcttggac 1561 tctgtgtctg tggactgggc accatccctg ctgagcacct gtcccggcgt cctaaaggag 1621 tatgttgtcc gctgccgaga tgaagacagc aaacaggtgt cagagcatcc cgtgcagccc 1681 acagagaccc aagttaccct cagtggcctg cgggctggtg tagcctacac ggtgcaggtg 1741 cgagcagaca cagcgtggct gaggggtgtc tggagccagc cccagcgctt cagcatcgaa 1801 gtgcaggttt ctgattggct catcttcttc gcctccctgg ggagcttcct gagcatcctt 1861 ctcgtgggcg tccttggcta ccttggcctg aacagggccg cacggcacct gtgcccgccg 1921 ctgcccacac cctgtgccag ctccgccatt gagttccctg gagggaagga gacttggcag 1981 tggatcaacc cagtggactt ccaggaagag gcatccctgc aggaggccct ggtggtagag 2041 atgtcctggg acaaaggcga gaggactgag cctctcgaga agacagagct acctgagggt 2101 gcccctgagc tggccctgga tacagagttg tccttggagg atggagacag atgtgatcgt 2161 tgaggctcag agagggtgag tgactcgccc gaggctacgt agcacacaca ggagtcacat 2221 ttggacccaa ataacccaga gctcctccag gctccagtgc acctgcctcc tctctgcccc 2281 gtgcctgttg ccacccatcc tgcgggggaa ccctagatgc tgccatgaaa tggaagctgc 2341 tgcaccctgc tgggcctggc atccgtgggg caggagcaga ccctgccatt tacctgttct 2401 ggcgtagaat ggactgggaa tgggggcaag gggggctcag atggatccct ggaccctggg 2461 ctgggcatcc acccccagga gcactggatg gggagtctgg actcaagggc tccctgcagc 2521 attgcggggt cttgtagctt ggaggatcca ggcatatagg gaagggggct gtaaactttg 2581 tgggaaaaat gacggtcctc ccatcccacc ccccacccca ccctcacccc cctataaaat 2641 gggggtggtg ataatgacct tacacagctg ttcaaaatca tcgtaaatga gcctcctctt 2701 gggtattttt ttcctgtttg aagcttgaat gtcctgctca aaatctcaaa acacgagcct 2761 tggaattcaa aaaaaaaaaa aaaaaaa Human IL-12R 131 mRNA Variant 4 (SEQ ID NO: 7) 1 agaacactcc gctgcctctc cagagccagg cacacagcag gcgctccata aatgttcgtt 61 ggtcttttct ccttgctcag cttcaatgtg ttccggagtg gggacggggt ggctgaacct 121 cgcaggtggc agagaggctc ccctggggct gtggggctct acgtggatcc gatggagccg 181 ctggtgacct gggtggtccc cctcctcttc ctcttcctgc tgtccaggca gggcgctgcc 241 tgcagaacca gtgagtgctg ttttcaggac ccgccatatc cggatgcaga ctcaggctcg 301 gcctcgggcc ctagggacct gagatgctat cggatatcca gtgatcgtta cgagtgctcc 361 tggcagtatg agggtcccac agctggggtc agccacttcc tgcggtgttg ccttagctcc 421 gggcgctgct gctacttcgc cgccggctca gccaccaggc tgcagttctc cgaccaggct 481 ggggtgtctg tgctgtacac tgtcacactc tgggtggaat cctgggccag gaaccagaca 541 gagaagtctc ctgaggtgac cctgcagctc tacaactcag ttaaatatga gcctcctctg 601 ggagacatca aggtgtccaa gttggccggg cagctgcgta tggagtggga gaccccggat 661 aaccaggttg gtgctgaggt gcagttccgg caccggacac ccagcagccc atggaagttg 721 ggcgactgcg gacctcagga tgatgatact gagtcctgcc tctgccccct ggagatgaat 781 gtggcccagg aattccagct ccgacgacgg cagctgggga gccaaggaag ttcctggagc 841 aagtggagca gccccgtgtg cgttccccct gaaaaccccc cacagcctca ggtgagattc 901 tcggtggagc agctgggcca ggatgggagg aggcggctga ccctgaaaga gcagccaacc 961 cagctggagc ttccagaagg ctgtcaaggg ctggcgcctg gcacggaggt cacttaccga 1021 ctacagctcc acatgctgtc ctgcccgtgt aaggccaagg ccaccaggac cctgcacctg 1081 gggaagatgc cctatctctc gggtgctgcc tacaacgtgg ctgtcatctc ctcgaaccaa 1141 tttggtcctg gcctgaacca gacgtggcac attcctgccg acacccacac agaaccagtg 1201 gctctgaata tcagcgtcgg aaccaacggg accaccatgt attggccagc ccgggctcag 1261 agcatgacgt attgcattga atggcagcct gtgggccagg acgggggcct tgccacctgc 1321 agcctgactg cgccgcaaga cccggatccg gctggaatgg caacctacag ctggagtcga 1381 gagtctgggg caatggggca ggaaaagtgt tactacatta ccatctttgc ctctgcgcac 1441 cccgagaagc tcaccttgtg gtctacggtc ctgtccacct accactttgg gggcaatgcc 1501 tcagcagctg ggacaccgca ccacgtctcg gtgaagaatc atagcttgga ctctgtgtct 1561 gtggactggg caccatccct gctgagcacc tgtcccggcg tcctaaagga gtatgagtc 1621 cgctgccgag atgaagacag caaacaggtg tcagagcatc ccgtgcagcc cacagagacc 1681 caagttaccc tcagtggcct gcgggctggt gtagcctaca cggtgcaggt gcgagcagac 1741 acagcgtggc tgaggggtgt ctggagccag ccccagcgct tcagcatcga agtgcaggtt 1801 tctgattggc tcatcactt cgcctccctg gggagcttcc tgagcatcct tctcgtgggc 1861 gtccttggct accttggcct gaacagggcc gcacggcacc tgtgcccgcc gctgcccaca 1921 ccctgtgcca gctccgccat tgagttccct ggagggaagg agacttggca gtggatcaac 1981 ccagtggact tccaggaaga ggcatccctg caggaggccc tggtggtaga gatgtcctgg 2041 gacaaaggcg agaggactga gcctctcgag aagacagagc tacctgaggg tgcccctgag 2101 ctggccctgg atacagagtt gtccttggag gatggagaca ggtgcaaggc caagatgtga 2161 tcgttgaggc tcagagaggg tgagtgactc gcccgaggct acgtagcaca cacaggagtc 2221 acatttggac ccaaataacc cagagctcct ccaggctcca gtgcacctgc ctcctctctg 2281 ccccgtgcct gttgccaccc atcctgcggg ggaaccctag atgctgccat gaaatggaag 2341 ctgctgcacc ctgctgggcc tggcatccgt ggggcaggag cagaccctgc catttacctg 2401 ttctggcgta gaatggactg ggaatggggg caaggggggc tcagatggat ccctggaccc 2461 tgggctgggc atccaccccc aggagcactg gatggggagt ctggactcaa gggctccctg 2521 cagcattgcg gggtcttgta gcttggagga tccaggcata tagggaaggg ggctgtaaac 2581 tagtgggaa aaatgacggt cctcccatcc caccccccac cccaccctca cccccctata 2641 aaatgggggt ggtgataatg accttacaca gctgttcaaa atcatcgtaa atgagcctcc 2701 tcttgggtat attacctg atgaagctt gaatgtcctg ctcaaaatct caaaacacga 2761 gccttggaat tcaaaaaaaa aaaaaaaaaa a Human IL-12R 132 mRNA Variant 1 (SEQ ID NO: 8) 1 tgcagagcac agagaaagga catctgcgag gaaagttccc tgatggctgt caacaaagtg 61 ccacgtctct atggctgtga acgctgagca cacgattaa tcgcgcctat catatcttgg 121 tgcataaacg cacctcacct cggtcaaccc ttgctccgtc ttatgagaca ggctttatta 181 tccgcatat atatgagggg aaactgacgg tggagagaga attatcttgc tcaaggcgac 241 acagcagagc ccacaggtgg cagaatccca cccgagcccg cttcgacccg cggggtggaa 301 accacgggcg cccgcccggc tgcgcttcca gagctgaact gagaagcgag tcctctccgc 361 cctgcggcca ccgcccagcc ccgacccccg ccccggcccg atcctcactc gccgccagct 421 ccccgcgccc accccggagt tggtggcgca gaggcgggag gcggaggcgg gagggcgggc 481 gctggcaccg ggaacgcccg agcgccggca gagagcgcgg agagcgcgac acgtgcggcc 541 cagagcaccg gggccacccg gtccccgcag gcccgggacc gcgcccgctg gcaggcgaca 601 cgtggaagaa tacggagttc tataccagag ttgattgag atggcacata cattagagg 661 atgctcattg gcatttatgt ttataatcac gtggctgttg attaaagcaa aaatagatgc 721 gtgcaagaga ggcgatgtga ctgtgaagcc ttcccatgta attttacttg gatccactgt 781 caatattaca tgctctttga agcccagaca aggctgcttt cactattcca gacgtaacaa 841 gttaatcctg tacaagtag acagaagaat caattacac catggccact ccctcaattc 901 tcaagtcaca ggtcttcccc ttggtacaac cagtagtc tgcaaactgg cctgtatcaa 961 tagtgatgaa attcaaatat gtggagcaga gatcacga ggtgttgctc cagaacagcc 1021 tcaaaattta tcctgcatac agaagggaga acaggggact gtggcctgca cctgggaaag 1081 aggacgagac acccacttat acactgagta tactctacag ctaagtggac caaaaaattt 1141 aacctggcag aagcaatgta aagacattta ttgtgactat ttggactttg gaatcaacct 1201 cacccctgaa tcacctgaat ccaatttcac agccaaggtt actgctgtca atagtcttgg 1261 aagctcctct tcacttccat ccacattcac attcttggac atagtgaggc ctcttcctcc 1321 gtgggacatt agaatcaaat ttcaaaaggc ttctgtgagc agatgtaccc tttattggag 1381 agatgaggga ctggtactgc ttaatcgact cagatatcgg cccagtaaca gcaggctctg 1441 gaatatggtt aatgttacaa aggccaaagg aagacatgat ttgctggatc tgaaaccatt 1501 tacagaatat gaatttcaga tttcctctaa gctacatctt tataagggaa gttggagtga 1561 ttggagtgaa tcattgagag cacaaacacc agaagaagag cctactggga tgttagatgt 1621 ctggtacatg aaacggcaca ttgactacag tagacaacag atttctcttt tctggaagaa 1681 tctgagtgtc tcagaggcaa gaggaaaaat tctccactat caggtgacct tgcaggagct 1741 gacaggaggg aaagccatga cacagaacat cacaggacac acctcctgga ccacagtcat 1801 tcctagaacc ggaaattggg ctgtggctgt gtctgcagca aattcaaaag gcagttctct 1861 gcccactcgt attaacataa tgaacctgtg tgaggcaggg ttgctggctc ctcgccaggt 1921 ctctgcaaac tcagagggca tggacaacat tctggtgact tggcagcctc ccaggaaaga 1981 tccctctgct gttcaggagt acgtggtgga atggagagag ctccatccag ggggtgacac 2041 acaggtccct ctaaactggc tacggagtcg accctacaat gtgtctgctc tgatttcaga 2101 gaacataaaa tcctacatct gttatgaaat ccgtgtgtat gcactctcag gggatcaagg 2161 aggatgcagc tccatcctgg gtaactctaa gcacaaagca ccactgagtg gcccccacat 2221 taatgccatc acagaggaaa aggggagcat tttaatttca tggaacagca ttccagtcca 2281 ggagcaaatg ggctgcctcc tccattatag gatatactgg aaggaacggg actccaactc 2341 ccagcctcag ctctgtgaaa ttccctacag agtctcccaa aattcacatc caataaacag 2401 cctgcagccc cgagtgacat atgtcctgtg gatgacagct ctgacagctg ctggtgaaag 2461 ttcccacgga aatgagaggg aattttgtct gcaaggtaaa gccaattgga tggcgtttgt 2521 ggcaccaagc atttgcattg ctatcatcat ggtgggcatt ttctcaacgc attacttcca 2581 gcaaaaggtg tttgttctcc tagcagccct cagacctcag tggtgtagca gagaaattcc 2641 agatccagca aatagcactt gcgctaagaa atatcccatt gcagaggaga agacacagct 2701 gcccttggac aggctcctga tagactggcc cacgcctgaa gatcctgaac cgctggtcat 2761 cagtgaagtc cttcatcaag tgaccccagt tttcagacat cccccctgct ccaactggcc 2821 acaaagggaa aaaggaatcc aaggtcatca ggcctctgag aaagacatga tgcacagtgc 2881 ctcaagccca ccacctccaa gagctctcca agctgagagc agacaactgg tggatctgta 2941 caaggtgctg gagagcaggg gctccgaccc aaagcccgaa aacccagcct gtccctggac 3001 ggtgctccca gcaggtgacc ttcccaccca tgatggctac ttaccctcca acatagatga 3061 cctcccctca catgaggcac ctctcgctga ctctctggaa gaactggagc ctcagcacat 3121 ctccctttct gttttcccct caagttctct tcacccactc accttctcct gtggtgataa 3181 gctgactctg gatcagttaa agatgaggtg tgactccctc atgctctgag tggtgaggct 3241 tcaagcctta aagtcagtgt gccctcaacc agcacagcct gccccaattc ccccagcccc 3301 tgctccagca gctgtcatct ctgggtgcca ccatcggtct ggctgcagct agaggacagg 3361 caagccagct ctgggggagt cttaggaact gggagttggt cttcactcag atgcctcatc 3421 ttgcctttcc cagggcctta aaattacatc cttcactgtg tggacctaga gactccaact 3481 tgaattccta gtaactttct tggtatgctg gccagaaagg gaaatgagga ggagagtaga 3541 aaccacagct cttagtagta atggcataca gtctagagga ccattcatgc aatgactatt 3601 tctaaagcac ctgctacaca gcaggctgta cacagcagat cagtactgtt caacagaact 3661 tcctgagatg atggaaatgt tctacctctg cactcactgt ccagtacatt agacactagg 3721 cacattggct gttaatcact tggaatgtgt ttagcttgac tgaggaatta aattttgatt 3781 gtaaatttaa atcgccacac atggctagtg gctactgtat tggagtgcac agctctagat 3841 ggctcctaga ttattgagag ccttcaaaac aaatcaacct agttctatag atgaagacat 3901 aaaagacact ggtaaacacc aaggtaaaag ggcccccaag gtggtcatga ctggtctcat 3961 ttgcagaagt ctaagaatgt acctttttct ggccgggcgt ggtagctcat gcctgtaatc 4021 ccagcacttt gggaggctga Human IL-12R 132 mRNA Variant 2 (SEQ ID NO: 9) 1 tgcagagcac agagaaagga catctgcgag gaaagttccc tgatggctgt caacaaagtg 61 ccacgtctct atggctgtga acgctgagca cacgatttta tcgcgcctat catatcttgg 121 tgcataaacg cacctcacct cggtcaaccc ttgctccgtc ttatgagaca ggctttatta 181 tccgcatttt atatgagggg aaactgacgg tggagagaga attatcttgc tcaaggcgac 241 acagcagagc ccacaggtgg cagaatccca cccgagcccg cttcgacccg cggggtggaa 301 accacgggcg cccgcccggc tgcgcttcca gagctgaact gagaagcgag tcctctccgc 361 cctgcggcca ccgcccagcc ccgacccccg ccccggcccg atcctcactc gccgccagct 421 ccccgcgccc accccggagt tggtggcgca gaggcgggag gcggaggcgg gagggcgggc 481 gctggcaccg ggaacgcccg agcgccggca gagagcgcgg agagcgcgac acgtgcggcc 541 cagagcaccg gggccacccg gtccccgcag gcccgggacc gcgcccgctg gcaggcgaca 601 cgtggtcacg gtgatccatt tgtaaagtcg ggaataaatg acctctgaag tgttgtctgt 661 atattgatct gctaccagta aaacatatct ctgaagaata cggagttcta taccagagtt 721 gattgttgat ggcacatact tttagaggat gctcattggc atttatgttt ataatcacgt 781 ggctgttgat taaagcaaaa atagatgcgt gcaagagagg cgatgtgact gtgaagcctt 841 cccatgtaat tttacttgga tccactgtca atattacatg ctctttgaag cccagacaag 901 gctgctttca ctattccaga cgtaacaagt taatcctgta caagtttgac agaagaatca 961 attttcacca tggccactcc ctcaattctc aagtcacagg tcttcccctt ggtacaacct 1021 tgtttgtctg caaactggcc tgtatcaata gtgatgaaat tcaaatatgt ggagcagaga 1081 tcttcgttgg tgttgctcca gaacagcctc aaaatttatc ctgcatacag aagggagaac 1141 aggggactgt ggcctgcacc tgggaaagag gacgagacac ccacttatac actgagtata 1201 ctctacagct aagtggacca aaaaatttaa cctggcagaa gcaatgtaaa gacatttatt 1261 gtgactattt ggactttgga atcaacctca cccctgaatc acctgaatcc aatttcacag 1321 ccaaggttac tgctgtcaat agtcttggaa gctcctcttc acttccatcc acattcacat 1381 tcttggacat agtgaggcct cttcctccgt gggacattag aatcaaattt caaaaggctt 1441 ctgtgagcag atgtaccctt tattggagag atgagggact ggtactgctt aatcgactca 1501 gatatcggcc cagtaacagc aggctctgga atatggttaa tgttacaaag gccaaaggaa 1561 gacatgattt gctggatctg aaaccattta cagaatatga atttcagatt tcctctaagc 1621 tacatcttta taagggaagt tggagtgatt ggagtgaatc attgagagca caaacaccag 1681 aagaagagcc tactgggatg ttagatgtct ggtacatgaa acggcacatt gactacagta 1741 gacaacagat ttctcttttc tggaagaatc tgagtgtctc agaggcaaga ggaaaaattc 1801 tccactatca ggtgaccttg caggagctga caggagggaa agccatgaca cagaacatca 1861 caggacacac ctcctggacc acagtcattc ctagaaccgg aaattgggct gtggctgtgt 1921 ctgcagcaaa ttcaaaaggc agttctctgc ccactcgtat taacataatg aacctgtgtg 1981 aggcagggtt gctggctcct cgccaggtct ctgcaaactc agagggcatg gacaacattc 2041 tggtgacttg gcagcctccc aggaaagatc cctctgctgt tcaggagtac gtggtggaat 2101 ggagagagct ccatccaggg ggtgacacac aggtccctct aaactggcta cggagtcgac 2161 cctacaatgt gtctgctctg atttcagaga acataaaatc ctacatctgt tatgaaatcc 2221 gtgtgtatgc actctcaggg gatcaaggag gatgcagctc catcctgggt aactctaagc 2281 acaaagcacc actgagtggc ccccacatta atgccatcac agaggaaaag gggagcattt 2341 taatttcatg gaacagcatt ccagtccagg agcaaatggg ctgcctcctc cattatagga 2401 tatactggaa ggaacgggac tccaactccc agcctcagct ctgtgaaatt ccctacagag 2461 tctcccaaaa ttcacatcca ataaacagcc tgcagccccg agtgacatat gtcctgtgga 2521 tgacagctct gacagctgct ggtgaaagtt cccacggaaa tgagagggaa ttagtctgc 2581 aaggtaaagc caattggatg gcgtttgtgg caccaagcat ttgcattgct atcatcatgg 2641 tgggcatta ctcaacgcat tacttccagc aaaagagaag acacagctgc ccttggacag 2701 gctcctgata gactggccca cgcctgaaga tcctgaaccg ctggtcatca gtgaagtcct 2761 tcatcaagtg accccagat tcagacatcc cccctgctcc aactggccac aaagggaaaa 2821 aggaatccaa ggtcatcagg cctctgagaa agacatgatg cacagtgcct caagcccacc 2881 acctccaaga gctctccaag ctgagagcag acaactggtg gatctgtaca aggtgctgga 2941 gagcaggggc tccgacccaa agcccgaaaa cccagcctgt ccctggacgg tgctcccagc 3001 aggtgacctt cccacccatg atggctactt accctccaac atagatgacc tcccctcaca 3061 tgaggcacct ctcgctgact ctctggaaga actggagcct cagcacatct ccctactgt 3121 tacccctca agactcttc acccactcac cttctcctgt ggtgataagc tgactctgga 3181 tcagttaaag atgaggtgtg actccctcat gctctgagtg gtgaggcttc aagccttaaa 3241 gtcagtgtgc cctcaaccag cacagcctgc cccaattccc ccagcccctg ctccagcagc 3301 tgtcatctct gggtgccacc atcggtctgg ctgcagctag aggacaggca agccagctct 3361 gggggagtct taggaactgg gagttggtct tcactcagat gcctcatctt gccatccca 3421 gggccttaaa attacatcct tcactgtgtg gacctagaga ctccaacttg aattcctagt 3481 aactacttg gtatgctggc cagaaaggga aatgaggagg agagtagaaa ccacagctct 3541 tagtagtaat ggcatacagt ctagaggacc attcatgcaa tgactatttc taaagcacct 3601 gctacacagc aggctgtaca cagcagatca gtactgttca acagaacttc ctgagatgat 3661 ggaaatgttc tacctctgca ctcactgtcc agtacattag acactaggca cattggctgt 3721 taatcacttg gaatgtgat agcttgactg aggaattaaa attgattgt aaatttaaat 3781 cgccacacat ggctagtggc tactgtattg gagtgcacag ctctagatgg ctcctagatt 3841 attgagagcc ttcaaaacaa atcaacctag ttctatagat gaagacataa aagacactgg 3901 taaacaccaa ggtaaaaggg cccccaaggt ggtcatgact ggtctcattt gcagaagtct 3961 aagaatgtac catactgg ccgggcgtgg tagctcatgc ctgtaatccc agcactttgg 4021 gaggctga Human IL-12R D2 mRNA Variant 3 (SEQ ID NO: 10) 1 tgcagagcac agagaaagga catctgcgag gaaagttccc tgatggctgt caacaaagtg 61 ccacgtctct atggctgtga acgctgagca cacgattaa tcgcgcctat catatcttgg 121 tgcataaacg cacctcacct cggtcaaccc ttgctccgtc ttatgagaca ggctttatta 181 tccgcatal atatgagggg aaactgacgg tggagagaga attatcttgc tcaaggcgac 241 acagcagagc ccacaggtgg cagaatccca cccgagcccg cttcgacccg cggggtggaa 301 accacgggcg cccgcccggc tgcgcttcca gagctgaact gagaagcgag tcctctccgc 361 cctgcggcca ccgcccagcc ccgacccccg ccccggcccg atcctcactc gccgccagct 421 ccccgcgccc accccggagt tggtggcgca gaggcgggag gcggaggcgg gagggcgggc 481 gctggcaccg ggaacgcccg agcgccggca gagagcgcgg agagcgcgac acgtgcggcc 541 cagagcaccg gggccacccg gtccccgcag gcccgggacc gcgcccgctg gcaggcgaca 601 cgtggaagaa tacggagttc tataccagag ttgattgttg atggcacata cttttagagg 661 atgctcattg gcatttatgt ttataatcac gtggctgttg attaaagcaa aaatagatgc 721 gtgcaagaga ggcgatgtga ctgtgaagcc ttcccatgta attttacttg gatccactgt 781 caatattaca tgctctttga agcccagaca aggctgcttt cactattcca gacgtaacaa 841 gttaatcctg tacaagtttg acagaagaat caattttcac catggccact ccctcaattc 901 tcaagtcaca ggtcttcccc ttggtacaac cttgtttgtc tgcaaactgg cctgtatcaa 961 tagtgatgaa attcaaatat gtggagcaga gatcttcgtt ggtgttgctc cagaacagcc 1021 tcaaaattta tcctgcatac agaagggaga acaggggact gtggcctgca cctgggaaag 1081 aggacgagac acccacttat acactgagta tactctacag ctaagtggac caaaaaattt 1141 aacctggcag aagcaatgta aagacattta ttgtgactat ttggactttg gaatcaacct 1201 cacccctgaa tcacctgaat ccaatttcac agccaaggtt actgctgtca atagtcttgg 1261 aagctcctct tcacttccat ccacattcac attcttggac atagtgaggc ctcttcctcc 1321 gtgggacatt agaatcaaat ttcaaaaggc ttctgtgagc agatgtaccc tttattggag 1381 agatgaggga ctggtactgc ttaatcgact cagatatcgg cccagtaaca gcaggctctg 1441 gaatatggtt aatgttacaa aggccaaagg aagacatgat ttgctggatc tgaaaccatt 1501 tacagaatat gaatttcaga tttcctctaa gctacatctt tataagggaa gttggagtga 1561 ttggagtgaa tcattgagag cacaaacacc agaagaagag cctactggga tgttagatgt 1621 ctggtacatg aaacggcaca ttgactacag tagacaacag atttctcttt tctggaagaa 1681 tctgagtgtc tcagaggcaa gaggaaaaat tctccactat caggtgacct tgcaggagct 1741 gacaggaggg aaagccatga cacagaacat cacaggacac acctcctgga ccacagtcat 1801 tcctagaacc ggaaattggg ctgtggctgt gtctgcagca aattcaaaag gcagttctct 1861 gcccactcgt attaacataa tgaacctgtg tgaggcaggg ttgctggctc ctcgccaggt 1921 ctctgcaaac tcagagggca tggacaacat tctggtgact tggcagcctc ccaggaaaga 1981 tccctctgct gttcaggagt acgtggtgga atggagagag ctccatccag ggggtgacac 2041 acaggtccct ctaaactggc tacggagtcg accctacaat gtgtctgctc tgatttcaga 2101 aattccctac agagtctccc aaaattcaca tccaataaac agcctgcagc cccgagtgac 2161 atatgtcctg tggatgacag ctctgacagc tgctggtgaa agttcccacg gaaatgagag 2221 ggaattttgt ctgcaaggta aagccaattg gatggcgttt gtggcaccaa gcatttgcat 2281 tgctatcatc atggtgggca ttttctcaac gcattacttc cagcaaaagg tgtttgttct 2341 cctagcagcc ctcagacctc agtggtgtag cagagaaatt ccagatccag caaatagcac 2401 ttgcgctaag aaatatccca ttgcagagga gaagacacag ctgcccttgg acaggctcct 2461 gatagactgg cccacgcctg aagatcctga accgctggtc atcagtgaag tccttcatca 2521 agtgacccca gttttcagac atcccccctg ctccaactgg ccacaaaggg aaaaaggaat 2581 ccaaggtcat caggcctctg agaaagacat gatgcacagt gcctcaagcc caccacctcc 2641 aagagctctc caagctgaga gcagacaact ggtggatctg tacaaggtgc tggagagcag 2701 gggctccgac ccaaagcccg aaaacccagc ctgtccctgg acggtgctcc cagcaggtga 2761 ccttcccacc catgatggct acttaccctc caacatagat gacctcccct cacatgaggc 2821 acctctcgct gactctctgg aagaactgga gcctcagcac atctcccttt ctgttttccc 2881 ctcaagttct cttcacccac tcaccttctc ctgtggtgat aagctgactc tggatcagtt 2941 aaagatgagg tgtgactccc tcatgctctg agtggtgagg cttcaagcct taaagtcagt 3001 gtgccctcaa ccagcacagc ctgccccaat tcccccagcc cctgctccag cagctgtcat 3061 ctctgggtgc caccatcggt ctggctgcag ctagaggaca ggcaagccag ctctggggga 3121 gtcttaggaa ctgggagttg gtcttcactc agatgcctca tcttgccttt cccagggcct 3181 taaaattaca tccttcactg tgtggaccta gagactccaa cttgaattcc tagtaacttt 3241 cttggtatgc tggccagaaa gggaaatgag gaggagagta gaaaccacag ctcttagtag 3301 taatggcata cagtctagag gaccattcat gcaatgacta tttctaaagc acctgctaca 3361 cagcaggctg tacacagcag atcagtactg ttcaacagaa cttcctgaga tgatggaaat 3421 gttctacctc tgcactcact gtccagtaca ttagacacta ggcacattgg ctgttaatca 3481 cttggaatgt gtttagcttg actgaggaat taaattttga ttgtaaattt aaatcgccac 3541 acatggctag tggctactgt attggagtgc acagctctag atggctccta gattattgag 3601 agccttcaaa acaaatcaac ctagttctat agatgaagac ataaaagaca ctggtaaaca 3661 ccaaggtaaa agggccccca aggtggtcat gactggtctc atttgcagaa gtctaagaat 3721 gtaccttttt ctggccgggc gtggtagctc atgcctgtaa tcccagcact ttgggaggct 3781 ga Human IL-12R 132 mRNA Variant 4 (SEQ ID NO: 11) 1 tgcagagcac agagaaagga catctgcgag gaaagttccc tgatggctgt caacaaagtg 61 ccacgtctct atggctgtga acgctgagca cacgatttta tcgcgcctat catatcttgg 121 tgcataaacg cacctcacct cggtcaaccc ttgctccgtc ttatgagaca ggctttatta 181 tccgcatttt atatgagggg aaactgacgg tggagagaga attatcttgc tcaaggcgac 241 acagcagagc ccacaggtgg cagaatccca cccgagcccg cttcgacccg cggggtggaa 301 accacgggcg cccgcccggc tgcgcttcca gagctgaact gagaagcgag tcctctccgc 361 cctgcggcca ccgcccagcc ccgacccccg ccccggcccg atcctcactc gccgccagct 421 ccccgcgccc accccggagt tggtggcgca gaggcgggag gcggaggcgg gagggcgggc 481 gctggcaccg ggaacgcccg agcgccggca gagagcgcgg agagcgcgac acgtgcggcc 541 cagagcaccg gggccacccg gtccccgcag gcccgggacc gcgcccgctg gcaggcgaca 601 cgtggaagaa tacggagttc tataccagag ttgattgttg atggcacata cttttagagg 661 atgctcattg gcatttatgt ttataatcac gtggctgttg attaaagcaa aaatagatgc 721 gtgcaagaga ggcgatgtga ctgtgaagcc ttcccatgta attttacttg gatccactgt 781 caatattaca tgctctttga agcccagaca aggctgcttt cactattcca gacgtaacaa 841 gttaatcctg tacaagtttg acagaagaat caattttcac catggccact ccctcaattc 901 tcaagtcaca ggtcttcccc ttggtacaac cttgtttgtc tgcaaactgg cctgtatcaa 961 tagtgatgaa attcaaatat gtggagcaga gatcttcgtt ggtgttgctc cagaacagcc 1021 tcaaaattta tcctgcatac agaagggaga acaggggact gtggcctgca cctgggaaag 1081 aggacgagac acccacttat acactgagta tactctacag ctaagtggac caaaaaattt 1141 aacctggcag aagcaatgta aagacattta ttgtgactat ttggactttg gaatcaacct 1201 cacccctgaa tcacctgaat ccaatttcac agccaaggtt actgctgtca atagtcttgg 1261 aagctcctct tcacttccat ccacattcac attcttggac atagtgaggc ctcttcctcc 1321 gtgggacatt agaatcaaat ttcaaaaggc ttctgtgagc agatgtaccc tttattggag 1381 agatgaggga ctggtactgc ttaatcgact cagatatcgg cccagtaaca gcaggctctg 1441 gaatatggtt aatgttacaa aggccaaagg aagacatgat ttgctggatc tgaaaccatt 1501 tacagaatat gaatttcaga tttcctctaa gctacatctt tataagggaa gttggagtga 1561 ttggagtgaa tcattgagag cacaaacacc agaagaagag cctactggga tgttagatgt 1621 ctggtacatg aaacggcaca ttgactacag tagacaacag atttctcttt tctggaagaa 1681 tctgagtgtc tcagaggcaa gaggaaaaat tctccactat caggtgacct tgcaggagct 1741 gacaggaggg aaagccatga cacagaacat cacaggacac acctcctgga ccacagtcat 1801 tcctagaacc ggaaattggg ctgtggctgt gtctgcagca aattcaaaag gcagttctct 1861 gcccactcgt attaacataa tgaacctgtg tgaggcaggg ttgctggctc ctcgccaggt 1921 ctctgcaaac tcagagggca tggacaacat tctggtgact tggcagcctc ccaggaaaga 1981 tccctctgct gttcaggagt acgtggtgga atggagagag ctccatccag ggggtgacac 2041 acaggtccct ctaaactggc tacggagtcg accctacaat gtgtctgctc tgatttcaga 2101 gaacataaaa tcctacatct gttatgaaat ccgtgtgtat gcactctcag gggatcaagg 2161 aggatgcagc tccatcctgg gtaactctaa gcacaaagca ccactgagtg gcccccacat 2221 taatgccatc acagaggaaa aggggagcat tttaatttca tggaacagca ttccagtcca 2281 ggagcaaatg ggctgcctcc tccattatag gatatactgg aaggaacggg actccaactc 2341 ccagcctcag ctctgtgaaa ttccctacag agtctcccaa aattcacatc caataaacag 2401 cctgcagccc cgagtgacat atgtcctgtg gatgacagct ctgacagctg ctggtgaaag 2461 ttcccacgga aatgagaggg aattttgtct gcaaggagaa gacacagctg cccttggaca 2521 ggctcctgat agactggccc acgcctgaag atcctgaacc gctggtcatc agtgaagtcc 2581 ttcatcaagt gaccccagtt ttcagacatc ccccctgctc caactggcca caaagggaaa 2641 aaggaatcca aggtcatcag gcctctgaga aagacatgat gcacagtgcc tcaagcccac 2701 cacctccaag agctctccaa gctgagagca gacaactggt ggatctgtac aaggtgctgg 2761 agagcagggg ctccgaccca aagcccgaaa acccagcctg tccctggacg gtgctcccag 2821 caggtgacct tcccacccat gatggctact taccctccaa catagatgac ctcccctcac 2881 atgaggcacc tctcgctgac tctctggaag aactggagcc tcagcacatc tccctttctg 2941 ttttcccctc aagttctctt cacccactca ccttctcctg tggtgataag ctgactctgg 3001 atcagttaaa gatgaggtgt gactccctca tgctctgagt ggtgaggctt caagccttaa 3061 agtcagtgtg ccctcaacca gcacagcctg ccccaattcc cccagcccct gctccagcag 3121 ctgtcatctc tgggtgccac catcggtctg gctgcagcta gaggacaggc aagccagctc 3181 tgggggagtc ttaggaactg ggagttggtc ttcactcaga tgcctcatct tgcctttccc 3241 agggccttaa aattacatcc ttcactgtgt ggacctagag actccaactt gaattcctag 3301 taactttctt ggtatgctgg ccagaaaggg aaatgaggag gagagtagaa accacagctc 3361 ttagtagtaa tggcatacag tctagaggac cattcatgca atgactattt ctaaagcacc 3421 tgctacacag caggctgtac acagcagatc agtactgttc aacagaactt cctgagatga 3481 tggaaatgtt ctacctctgc actcactgtc cagtacatta gacactaggc acattggctg 3541 ttaatcactt ggaatgtgtt tagcttgact gaggaattaa attttgattg taaatttaaa 3601 tcgccacaca tggctagtgg ctactgtatt ggagtgcaca gctctagatg gctcctagat 3661 tattgagagc cttcaaaaca aatcaaccta gttctataga tgaagacata aaagacactg 3721 gtaaacacca aggtaaaagg gcccccaagg tggtcatgac tggtctcatt tgcagaagtc 3781 taagaatgta cctttttctg gccgggcgtg gtagctcatg cctgtaatcc cagcactttg 3841 ggaggctga Human IL-23R mRNA (SEQ ID NO: 12) 1 acaagggtgg cagcctggct ctgaagtgga attatgtgct tcaaacaggt tgaaagaggg 61 aaacagtctt ttcctgcttc cagacatgaa tcaggtcact attcaatggg atgcagtaat 121 agccctttac atactcttca gctggtgtca tggaggaatt acaaatataa actgctctgg 181 ccacatctgg gtagaaccag ccacaatttt taagatgggt atgaatatct ctatatattg 241 ccaagcagca attaagaact gccaaccaag gaaacttcat ttttataaaa atggcatcaa 301 agaaagattt caaatcacaa ggattaataa aacaacagct cggctttggt ataaaaactt 361 tctggaacca catgcttcta tgtactgcac tgctgaatgt cccaaacatt ttcaagagac 421 actgatatgt ggaaaagaca tttcttctgg atatccgcca gatattcctg atgaagtaac 481 ctgtgtcatt tatgaatatt caggcaacat gacttgcacc tggaatgctg ggaagctcac 541 ctacatagac acaaaatacg tggtacatgt gaagagttta gagacagaag aagagcaaca 601 gtatctcacc tcaagctata ttaacatctc cactgattca ttacaaggtg gcaagaagta 661 cttggtttgg gtccaagcag caaacgcact aggcatggaa gagtcaaaac aactgcaaat 721 tcacctggat gatatagtga taccttctgc agccgtcatt tccagggctg agactataaa 781 tgctacagtg cccaagacca taatttattg ggatagtcaa acaacaattg aaaaggtttc 841 ctgtgaaatg agatacaagg ctacaacaaa ccaaacttgg aatgttaaag aatttgacac 901 caattttaca tatgtgcaac agtcagaatt ctacttggag ccaaacatta agtacgtatt 961 tcaagtgaga tgtcaagaaa caggcaaaag gtactggcag ccttggagtt caccgttttt 1021 tcataaaaca cctgaaacag ttccccaggt cacatcaaaa gcattccaac atgacacatg 1081 gaattctggg ctaacagttg cttccatctc tacagggcac cttacttctg acaacagagg 1141 agacattgga cttttattgg gaatgatcgt ctttgctgtt atgttgtcaa ttctttcttt 1201 gattgggata tttaacagat cattccgaac tgggattaaa agaaggatct tattgttaat 1261 accaaagtgg ctttatgaag atattcctaa tatgaaaaac agcaatgttg tgaaaatgct 1321 acaggaaaat agtgaactta tgaataataa ttccagtgag caggtcctat atgttgatcc 1381 catgattaca gagataaaag aaatcttcat cccagaacac aagcctacag actacaagaa 1441 ggagaataca ggacccctgg agacaagaga ctacccgcaa aactcgctat tcgacaatac 1501 tacagttgta tatattcctg atctcaacac tggatataaa ccccaaattt caaattttct 1561 gcctgaggga agccatctca gcaataataa tgaaattact tccttaacac ttaaaccacc 1621 agttgattcc ttagactcag gaaataatcc caggttacaa aagcatccta attttgcttt 1681 ttctgtttca agtgtgaatt cactaagcaa cacaatattt cttggagaat taagcctcat 1741 attaaatcaa ggagaatgca gttctcctga catacaaaac tcagtagagg aggaaaccac 1801 catgcttttg gaaaatgatt cacccagtga aactattcca gaacagaccc tgcttcctga 1861 tgaatttgtc tcctgtttgg ggatcgtgaa tgaggagttg ccatctatta atacttattt 1921 tccacaaaat attttggaaa gccacttcaa taggatttca ctcttggaaa agtagagctg 1981 tgtggtcaaa atcaatatga gaaagctgcc ttgcaatctg aacttgggtt ttccctgcaa 2041 tagaaattga attctgcctc tttttgaaaa aaatgtattc acatacaaat cttcacatgg 2101 acacatgttt tcatttccct tggataaata cctaggtagg ggattgctgg gccatatgat 2161 aagcatatgt ttcagttcta ccaatcttgt ttccagagta gtgacatttc tgtgctccta 2221 ccatcaccat gtaagaattc ccgggagctc catgcctttt taattttagc cattcttctg 2281 cctcatttct taaaattaga gaattaaggt cccgaaggtg gaacatgctt catggtcaca 2341 catacaggca caaaaacagc attatgtgga cgcctcatgt attttttata gagtcaacta 2401 tttcctcttt attttccctc attgaaagat gcaaaacagc tctctattgt gtacagaaag 2461 ggtaaataat gcaaaatacc tggtagtaaa ataaatgctg aaaattttcc tttaaaatag 2521 aatcattagg ccaggcgtgg tggctcatgc ttgtaatccc agcactttgg taggctgagg 2581 tgggtggatc acctgaggtc aggagttcga gtccagcctg gccaatatgc tgaaaccctg 2641 tctctactaa aattacaaaa attagccggc catggtggca ggtgcttgta atcccagcta 2701 cttgggaggc tgaggcagga gaatcacttg aaccaggaag gcagaggttg cactgagctg 2761 agattgtgcc actgcactcc agcctgggca acaagagcaa aactctgtct ggaaaaaaaa 2821 aaaaaa An antisense nucleic acid molecule can be complementary to all or part of a non-coding region of the coding strand of a nucleotide sequence encoding an IL-12A
(p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R protein. Non-coding regions (5' and 3' untranslated regions) are the 5' and 3' sequences that flank the coding region in a gene and are not translated into amino acids.
Based upon the sequences disclosed herein, one of skill in the art can easily choose and synthesize any of a number of appropriate antisense nucleic acids to target a nucleic acid encoding an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R
protein described herein. Antisense nucleic acids targeting a nucleic acid encoding an IL-12A
(p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R protein can be designed using the software available at the Integrated DNA Technologies website.
An antisense nucleic acid can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides or more in length. An antisense oligonucleotide can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used.
Examples of modified nucleotides which can be used to generate an antisense nucleic acid include 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethy1-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethy1-2-thiouracil, beta-D-mannosylqueosine, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methy1-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methy1-2-thiouracil, 3-(3-amino-3-N-2- carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an anti sense orientation to a target nucleic acid of interest).
The antisense nucleic acid molecules described herein can be prepared in vitro and administered to a mammal, e.g., a human. Alternatively, they can be generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding an IL-12A
(p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R protein to thereby inhibit expression, e.g., by inhibiting transcription and/or translation. The hybridization can be by conventional nucleotide complementarities to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA duplexes, through specific interactions in the major groove of the double helix. The antisense nucleic acid molecules can be delivered to a mammalian cell using a vector (e.g., a lentivirus, a retrovirus, or an adenovirus vector).
An antisense nucleic acid can be an a-anomeric nucleic acid molecule. An a-anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual, 13-units, the strands run parallel to each other (Gaultier et al., Nucleic Acids Res. 15:6625-6641, 1987). The antisense nucleic acid can also comprise a 2'-0-methylribonucleotide (Inoue et al., Nucleic Acids Res. 15:6131-6148, 1987) or a chimeric RNA-DNA analog (Inoue et al., FEBS Lett. 215:327-330, 1987). Non-limiting examples of antisense nucleic acids are described in Vaknin-Dembinsky et al., I Immunol.
176(12): 7768-7774, 2006.
Another example of an inhibitory nucleic acid is a ribozyme that has specificity for a nucleic acid encoding an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R protein (e.g., specificity for an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R 131, IL-12R 132, or IL-23R mRNA, e.g., specificity for any one of SEQ ID NOs: 1-12).
Ribozymes are catalytic RNA molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes (described in Haselhoff and Gerlach, Nature 334:585-591, 1988)) can be used to catalytically cleave mRNA transcripts to thereby inhibit translation of the protein encoded by the mRNA. A
ribozyme having specificity for an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R mRNA can be designed based upon the nucleotide sequence of any of the IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, and IL-23R
mRNA
sequences disclosed herein. For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R mRNA (see, e.g., U.S. Patent. Nos. 4,987,071 and 5,116,742).
Alternatively, an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R
(32, or IL-23R
mRNA can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel et al., Science 261:1411-1418, 1993.
An inhibitor nucleic acid can also be a nucleic acid molecule that forms triple helical structures. For example, expression of an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R
(31, IL-12R (32, or IL-23R protein can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the gene encoding the IL-12A (p35), (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R protein (e.g., the promoter and/or enhancer, e.g., a sequence that is at least 1 kb, 2 kb, 3 kb, 4 kb, or 5 kb upstream of the transcription initiation start state) to form triple helical structures that prevent transcription of the gene in target cells. See generally Helene, Anticancer Drug Des. 6(6):569-84, 1991;
Helene, Ann. N.Y. Acad. Sci. 660:27-36, 1992; and Maher, Bioassays 14(12):807-15, 1992.
In various embodiments, inhibitory nucleic acids can be modified at the base moiety, sugar moiety, or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acids can be modified to generate peptide nucleic acids (see, e.g., Hyrup et al., Bioorganic Medicinal Chem. 4(1):5-23, 1996). Peptide nucleic acids (PNAs) are nucleic acid mimics, e.g., DNA mimics, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs allows for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols (see, e.g., Perry-O'Keefe et al., Proc.
Natl. Acad. Sci.
U.S.A. 93:14670-675, 1996). PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication.
PNAs can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA
chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras can be generated which may combine the advantageous properties of PNA
and DNA. Such chimeras allow DNA recognition enzymes, e.g., RNAse H and DNA
polymerases, to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation.
The synthesis of PNA-DNA chimeras can be performed as described in Finn et al., Nucleic Acids Res. 24:3357-63, 1996. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry and modified nucleoside analogs. Compounds such as 5'-(4-methoxytrityl)amino-5'-deoxy-thymidine phosphoramidite can be used as a link between the PNA and the 5' end of DNA
(Mag et al., Nucleic Acids Res. 17:5973-88, 1989). PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5' PNA segment and a 3' DNA
segment (Finn et al., Nucleic Acids Res. 24:3357-63, 1996). Alternatively, chimeric molecules can be synthesized with a 5' DNA segment and a 3' PNA segment (Peterser et al., Bioorganic Med.
Chem. Lett. 5:1119-11124, 1975).
In some embodiments, the inhibitory nucleic acids can include other appended groups such as peptides, or agents facilitating transport across the cell membrane (see, Letsinger et al., Proc. Natl. Acad. Sci. U.S.A. 86:6553-6556, 1989; Lemaitre et al., Proc.
Natl. Acad. Sci.
U.S.A. 84:648-652, 1989; and WO 88/09810). In addition, the inhibitory nucleic acids can be modified with hybridization-triggered cleavage agents (see, e.g., Krol et al., Bio/Techniques 6:958-976, 1988) or intercalating agents (see, e.g., Zon, Pharm. Res. 5:539-549, 1988). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent, etc.
Another means by which expression of an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R mRNA can be decreased in a mammalian cell is by RNA
interference (RNAi). RNAi is a process in which mRNA is degraded in host cells. To inhibit an mRNA, double-stranded RNA (dsRNA) corresponding to a portion of the gene to be silenced (e.g., a gene encoding an IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R protein) is introduced into a mammalian cell. The dsRNA is digested into 21-23 nucleotide-long duplexes called short interfering RNAs (or siRNAs), which bind to a nuclease complex to form what is known as the RNA-induced silencing complex (or RISC).
The RISC targets the homologous transcript by base pairing interactions between one of the siRNA strands and the endogenous mRNA. It then cleaves the mRNA about 12 nucleotides from the 3' terminus of the siRNA (see Sharp et al., Genes Dev. 15:485-490, 2001, and Hammond et al., Nature Rev. Gen. 2:110-119, 2001).
RNA-mediated gene silencing can be induced in a mammalian cell in many ways, e.g., by enforcing endogenous expression of RNA hairpins (see, Paddison et al., Proc. Natl.
Acad. Sci. U.S.A. 99:1443-1448, 2002) or, as noted above, by transfection of small (21-23 nt) dsRNA (reviewed in Caplen, Trends Biotech. 20:49-51, 2002). Methods for modulating gene expression with RNAi are described, e.g., in U.S. Patent No. 6,506,559 and US
2003/0056235, which are hereby incorporated by reference.
Standard molecular biology techniques can be used to generate siRNAs. Short interfering RNAs can be chemically synthesized, recombinantly produced, e.g., by expressing RNA from a template DNA, such as a plasmid, or obtained from commercial vendors, such as Dharmacon. The RNA used to mediate RNAi can include synthetic or modified nucleotides, such as phosphorothioate nucleotides. Methods of transfecting cells with siRNA
or with plasmids engineered to make siRNA are routine in the art.
The siRNA molecules used to decrease expression of an IL-12A (p35), IL-12B
(p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R mRNA can vary in a number of ways. For example, they can include a 3' hydroxyl group and strands of 21, 22, or 23 consecutive nucleotides. They can be blunt ended or include an overhanging end at either the 3' end, the 5' end, or both ends. For example, at least one strand of the RNA molecule can have a 3' overhang from about 1 to about 6 nucleotides (e.g., 1-5, 1-3, 2-4 or 3-5 nucleotides (whether pyrimidine or purine nucleotides) in length. Where both strands include an overhang, the length of the overhangs may be the same or different for each strand.
To further enhance the stability of the RNA duplexes, the 3' overhangs can be stabilized against degradation (by, e.g., including purine nucleotides, such as adenosine or guanosine nucleotides or replacing pyrimidine nucleotides by modified analogues (e.g., substitution of uridine 2-nucleotide 3' overhangs by 2'-deoxythymidine is tolerated and does not affect the efficiency of RNAi). Any siRNA can be used in the methods of decreasing IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R mRNA, provided it has sufficient homology to the target of interest (e.g., a sequence present in any one of SEQ
ID NOs: 1-12, e.g., a target sequence encompassing the translation start site or the first exon of the mRNA). There is no upper limit on the length of the siRNA that can be used (e.g., the siRNA can range from about 21 base pairs of the gene to the full length of the gene or more (e.g., about 20 to about 30 base pairs, about 50 to about 60 base pairs, about 60 to about 70 base pairs, about 70 to about 80 base pairs, about 80 to about 90 base pairs, or about 90 to about 100 base pairs).
Non-limiting examples of siRNAs targeting IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R are described in Tan et al., I Alzheimers Dis. 38(3): 633-646, 2014; Niimi et al., I Neuroimmunol. 254(1-2):39-45, 2013. Non-limiting examples of short hairpin RNA (shRNA) targeting IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R are described in Bak et al., BMC Dermatol. 11:5, 2011.
Non-limiting examples of inhibitory nucleic acids are microRNAs (e.g., microRNA-29 (Brain et al., Immunity 39(3):521-536, 2013), miR-10a (Xue et al., I
Immunol.
187(11):5879-5886, 2011), microRNA-155 (Podsiad et al., Am. I Physiol. Lung Cell Mol.
Physiol. 310(5):L465-75, 2016).
In some embodiments, a therapeutically effective amount of an inhibitory nucleic acid targeting IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R can be administered to a subject (e.g., a human subject) in need thereof.
In some embodiments, the inhibitory nucleic acid can be about 10 nucleotides to about 40 nucleotides (e.g., about 10 to about 30 nucleotides, about 10 to about 25 nucleotides, about 10 to about 20 nucleotides, about 10 to about 15 nucleotides, 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotides, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36 nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, or 40 nucleotides) in length. One skilled in the art will appreciate that inhibitory nucleic acids may comprise at least one modified nucleic acid at either the 5' or 3' end of DNA or RNA.
Any of the inhibitor nucleic acids described herein can be formulated for administration to the gastrointestinal tract. See, e.g., the formulation methods described in US 2016/0090598 and Schoellhammer et al., Gastroenterology, doi:

10.1053/j.gastro.2017.01.002, 2017.
As is known in the art, the term "thermal melting point (Tm)" refers to the temperature, under defined ionic strength, pH, and inhibitory nucleic acid concentration, at which 50% of the inhibitory nucleic acids complementary to the target sequence hybridize to the target sequence at equilibrium. In some embodiments, an inhibitory nucleic acid can bind specifically to a target nucleic acid under stingent conditions, e.g., those in which the salt concentration is at least about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30 C. for short oligonucleotides (e.g., 10 to 50 nucleotide). Stringent conditions can also be achieved with the addition of destabilizing agents such as formamide.
In some embodiments of any of the inhibitory nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding any one of IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R) with a Tm of greater than 20 C, greater than 22 C, greater than 24 C, greater than 26 C, greater than 28 C, greater than 30 C, greater than 32 C, greater than 34 C, greater than 36 C, greater than 38 C, greater than 40 C, greater than 42 C, greater than 44 C, greater than 46 C, greater than 48 C, greater than 50 C, greater than 52 C, greater than 54 C, greater than 56 C, greater than 58 C, greater than 60 C, greater than 62 C, greater than 64 C, greater than 66 C, greater than 68 C, greater than 70 C, greater than 72 C, greater than 74 C, greater than 76 C, greater than 78 C, or greater than 80 C, e.g., as measured in phosphate buffered saline using a UV spectrophotometer.
In some embodiments of any of the inhibitor nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding any one of IL-12A (p35), IL-12B (p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R) with a Tm of about 20 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, about 30 C, about 28 C, about 26 C, about 24 C, or about 22 C (inclusive); about 22 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, about 30 C, about 28 C, about 26 C, or about (inclusive); about 24 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, about 30 C, about 28 C, or about 26 C (inclusive); about 26 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, about 30 C, or about 28 C (inclusive); about 28 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, or about 30 C (inclusive); about 30 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, or about 32 C (inclusive); about 32 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, or about 34 C (inclusive); about 34 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, or about 36 C
(inclusive); about 36 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, or about 38 C (inclusive); about 38 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, or about 40 C (inclusive); about 40 C
to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, or about 42 C
(inclusive); about 42 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, or about 44 C
(inclusive); about 44 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 .. C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, or about 46 C (inclusive); about 46 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, or about 48 C (inclusive); about 48 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, or about 50 C
(inclusive); about 50 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, or about 52 C (inclusive); about 52 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, or about 54 C
(inclusive); about 54 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, or about (inclusive); about 56 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, or about 58 C
(inclusive); about 58 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, or about 60 C (inclusive);
about 60 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, or about 62 C (inclusive); about 62 C
to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, or about 64 C (inclusive); about 64 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, or about 66 C (inclusive);
about 66 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, or about 68 C
(inclusive); about 68 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, or about 70 C (inclusive); about 70 C to about 80 C, about 78 C, about 76 C, about 74 C, or about 72 C (inclusive); about 72 C to about 80 C, about 78 C, about 76 C, or about 74 C (inclusive); about 74 C to about 80 C, about 78 C, or about 76 C (inclusive);
about 76 C to about 80 C or about 78 C (inclusive); or about 78 C to about (inclusive), In some embodiments, the inhibitory nucleic acid can be formulated in a nanoparticle (e.g., a nanoparticle including one or more synthetic polymers, e.g., Patil et al., Pharmaceutical Nanotechnol. 367:195-203, 2009; Yang et al., ACS Appl. Mater.
Interfaces, doi: 10.1021/acsami.6b16556, 2017; Perepelyuk et al., Mol. Ther. Nucleic Acids 6:259-268, 2017). In some embodiments, the nanoparticle can be a mucoadhesive particle (e.g., nanoparticles having a positively-charged exterior surface) (Andersen et al., Methods Mol.
Biol. 555:77-86, 2009). In some embodiments, the nanoparticle can have a neutrally-charged exterior surface.
In some embodiments, the inhibitory nucleic acid can be formulated, e.g., as a liposome (Buyens et al., I Control Release 158(3): 362-370, 2012; Scarabel et al., Expert Op/n. Drug Deliv. 17:1-14, 2017), a micelle (e.g., a mixed micelle) (Tangsangasaksri et al., BioMacromolecules 17:246-255, 2016; Wu et al., Nanotechnology, doi:
10.1088/1361-6528/aa6519, 2017), a microemulsion (WO 11/004395), a nanoemulsion, or a solid lipid nanoparticle (Sahay et al., Nature Biotechnol. 31:653-658, 2013; and Lin et al., Nanomedicine 9(1):105-120, 2014). Additional exemplary structural features of inhibitory nucleic acids and formulations of inhibitory nucleic acids are described in US
2016/0090598.
In some embodiments, a pharmaceutical composition can include a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In some examples, a pharmaceutical composition consists of a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In certain embodiments, the sterile saline is a pharmaceutical grade saline. In certain embodiments, a pharmaceutical composition can include one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition includes one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and phosphate-buffered saline (PBS). In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) and sterile phosphate-buffered saline (PBS). In some examples, the sterile saline is a pharmaceutical grade PBS.
In certain embodiments, one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) may be admixed with pharmaceutically acceptable .. active and/or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions depend on a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.
Pharmaceutical compositions including one or more inhibitory nucleic acids encompass any pharmaceutically acceptable salts, esters, or salts of such esters. Non-limiting examples of pharmaceutical compositions include pharmaceutically acceptable salts of inhibitory nucleic acids. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.
Also provided herein are prodrugs that can include additional nucleosides at one or both ends of an inhibitory nucleic acid which are cleaved by endogenous nucleases within the body, to form the active inhibitory nucleic acid.
Lipid moieties can be used to formulate an inhibitory nucleic acid. In certain such methods, the inhibitory nucleic acid is introduced into preformed liposomes or lipoplexes made of mixtures of cationic lipids and neutral lipids. In certain methods, inhibitory nucleic acid complexes with mono- or poly-cationic lipids are formed without the presence of a neutral lipid. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to a particular cell or tissue in a mammal. In some examples, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to fat tissue in a mammal. In certain embodiments, a lipid moiety is selected to increase distribution of an .. inhibitory nucleic acid to muscle tissue.
In certain embodiments, pharmaceutical compositions provided herein comprise one or more inhibitory nucleic acid and one or more excipients. In certain such embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose and polyvinylpyrrolidone.
In some examples, a pharmaceutical composition provided herein includes liposomes and emulsions. Liposomes and emulsions can be used to formulate hydrophobic compounds.
In some examples, certain organic solvents such as dimethylsulfoxide are used.
In some examples, a pharmaceutical composition provided herein includes one or more tissue-specific delivery molecules designed to deliver one or more inhibitory nucleic acids to specific tissues or cell types in a mammal. For example, a pharmaceutical composition can include liposomes coated with a tissue-specific antibody.
In some embodiments, a pharmaceutical composition provided herein can include a co-solvent system. Examples of such co-solvent systems include benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. A non-limiting example of such a co-solvent system is the VPD co-solvent system, which is a solution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80 and 65% w/v polyethylene glycol 300. As can be appreciated, other surfactants may be used instead of Polysorbate 80TM; the fraction size of polyethylene glycol may be varied;
other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone;
and other sugars or polysaccharides may substitute for dextrose.
In some examples, a pharmaceutical composition can be formulated for oral administration. In some examples, pharmaceutical compositions are formulated for buccal administration.
In some examples, a pharmaceutical composition is formulated for administration by injection (e.g., intravenous, subcutaneous, intramuscular, etc.). In some of these embodiments, a pharmaceutical composition includes a carrier and is formulated in aqueous solution, such as water or physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. In some examples, other ingredients are included (e.g., ingredients that aid in solubility or serve as preservatives).
In some examples, injectable suspensions are prepared using appropriate liquid carriers, suspending agents, and the like. Some pharmaceutical compositions for injection are formulated in unit dosage form, e.g., in ampoules or in multi-dose containers. Some pharmaceutical compositions for injection are suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, and/or dispersing agents.
Solvents suitable for use in pharmaceutical compositions for injection include, but are not limited to, lipophilic solvents and fatty oils, such as sesame oil, synthetic fatty acid esters, such as ethyl oleate or triglycerides, and liposomes.
Antibodies In some embodiments, the IL-12/IL-23 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to any one of IL-12A
(p35), IL-12B
(p40), IL-23 (p19), IL-12R (31, IL-12R (32, or IL-23R, or a combination thereof.
In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody .. can be a scFv-Fc, a VHEI domain, a VNAR domain, a (scFv)2, a minibody, or a BiTE. In some embodiments, an antibody can be a DVD-Ig, and a dual-affinity re-targeting antibody (DART), a triomab, kih IgG with a common LC, a crossmab, an ortho-Fab IgG, a 2-in-1-IgG, IgG-ScFv, seFv2-Fe, a bi-nanobody, tanden antibody, a DART-Fe, a scFv-HAS-scFv, DNL-Fab3, DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, .. knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, la-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH
IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody, nanobody-HSA, a diabody, a TandAb, scDiabody, scDiabody-CH3, Diabody-CH3, Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab')2-seFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody, dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HAS, tandem scFv, IgG-IgG, Cov-X-Body, and seFvl-PEG-seFv2.
Non-limiting examples of an antigen-binding fragment of an antibody include an Fv .. fragment, a Fab fragment, a F(ab')2 fragment, and a Fab' fragment.
Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgGl, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgGl, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgAl or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgAl or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).
In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Patent Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895;
US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US
.. 2017/0002082, each of which is incorporated by reference in its entirety.
In some embodiments, the antibody is ustekinumab (CNTO 1275, Stelarag) or a variant thereof (Krueger et al., N. Engl. I Med. 356(6):580-592, 2007;
Kauffman et al., Invest. Dermatol. 123(6):1037-1044, 2004; Gottlieb et al., Curr. Med. Res.
Op/n. 23(5):1081-1092, 2007; Leonardi et al., Lancet 371(9625):1665-1674, 2008; Papp et al., Lancet 371(9625):1675-1684, 2008). In some embodiments, the antibody is briakinumab (ABT-874, J-695) or a variant thereof (Gordon et al., I Invest. Dermatol. 132(2):304-314, 2012; Kimball et al., Arch Dermatol. 144(2): 200-207, 2008).
In some embodiments, the antibody is guselkumab (CNTO-1959) (Callis-Duffin et al., Am. Acad. Dermatol. 70(5 Suppl 1), 2014); AB162 (Sofen et al., I Allergy Cl/n.
Immunol.
133: 1032-40, 2014); tildrakizumab (MK-3222, 5CH900222) (Papp et al. (2015) Br.
Dermatol. 2015); Langley et al., Oral Presentation at: American Academy of Dermatology, March 21-25, Denver CO, 2014); AMG 139 (MEDI2070, brazikumab) (Gomollon, Gastroenterol. Hepatol. 38(Suppl. 1):13-19, 2015; Kock et al., Br. I
Pharmacol. 172(1):159-172, 2015); FM-202 (Tang et al., Immunology 135(2):112-124, 2012); FM-303 (Tang et al., Immunology 135(2):112-124, 2012); ADC-1012 (Tang et al., Immunology 135(2):112-124, 2012); LY-2525623 (Gaffen et al., Nat. Rev. Immunol. 14:585-600, 2014; Sands, Gastroenterol. Hepatol. 12(12):784-786, 2016), LY-3074828 (Coskun et al., Trends Pharmacol. Sci. 38(2):127-142, 2017), BI-655066 (risankizumab) (Singh et al., MAbs 7(4):778-791, 2015; Krueger et al., I Allergy Cl/n. Immunol. 136(1):116-124, 2015) or a variant thereof.
See e.g., Tang et al., Immunology 135(2):112-124, 2012. Further teachings of IL-12/IL-23 antibodies and antigen-binding fragments thereof are described in U.S. Patent Nos.

6,902,734; 7,247,711; 7,252,971; and 7,491,391; US 2012/0288494; and US
2013/0302343, each of which is incorporated by reference in its entirety.
In some embodiments, the IL-12/IL-23 inhibitor is PTG-200, an IL-23R inhibitor currently in preclinical development by Protagonist Therapeutics.
In some embodiments, the IL-12/IL-23 inhibitor is Mirikizumab (LY 3074828), an IL-23R inhibitor currently in clinical development (Phase II) by Eli Lilly.In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (KD) of less than 1 x 10-5M (e.g., less than 0.5 x 10-5M, less than 1 x 10' M, less than 0.5 x 10' M, less than 1 x 107M, less than 0.5 x 10-7M, less than 1 x 10-8M, less than 0.5 x 10-8M, less than 1 x 10-9M, less than 0.5 x 10-9M, less than 1 x 10' M, less than 0.5 x 10-1 M, less than 1 x 10-11M, less than 0.5 x 10-11M, or less than 1 x 10-12M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a KD of about lx 10-12M to about lx 10-5M, about 0.5 x 10-5M, about lx 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8 M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-1 M, about 0.5 x 10-1 M, about 1 x 10-11M, or about 0.5 x 10-11M (inclusive); about 0.5 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-1 M, about 0.5 x 10-1 M, or about 1 x 10-11M (inclusive); about 1 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-1 M, or about 0.5 x 10-1 M (inclusive); about 0.5 x 10-10 M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, or about 1 x 10-1 M
(inclusive);
about 1 x 10-1 M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9 M, or about 0.5 x 10-9M (inclusive); about 0.5 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5x 10-8M, or about 1 x 10-9M (inclusive); about lx 10-9M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, about 1 x 10-8M, or about 0.5 x 10-8M (inclusive); about 0.5 x 10-8M to about 1 x 10-5 M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 107M, about 0.5 x 10-7 M, or about 1 x 10-8 M (inclusive); about 1 x 10-8 M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 107M, or about 0.5 x 10' M
(inclusive);
about 0.5 x 10' M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, or about 1 x 107M (inclusive); about 1 x 10' M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, or about 0.5 x 10' M (inclusive); about 0.5 x 10' M to about 1 x 10-5M, about 0.5 x 10-5 M, or about 1 x 10' M (inclusive); about 1 x 10' M to about 1 x 10-5M or about 0.5 x 10-5 M (inclusive); or about 0.5 x 10-5M to about 1 x 10-5M
(inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Koff-of about lx 10' s-1 to about lx 10-3 s-1, about 0.5 x 10-3 s-1, about lx 10-4 s"
1, about 0.5 x 10-4 s-1, about 1 x 10-5 s-1, or about 0.5 x 10-5 s-1 (inclusive); about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, about 0.5 x 10-4 s-1, or about 1 x 10-5 s-1 (inclusive); about 1 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, or about 0.5 x 10-4 s-1 (inclusive); about 0.5 x 10-4 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, or about 1 x 10-4 s-1 (inclusive); about 1 x 10-4 s-lto about 1 x 10-3 s-1, or about 0.5 x 10-3 s-1 (inclusive); or about 0.5 x 10-5 s-lto about 1 x 10-3 s-1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Kon of about lx 102 M's' to about lx 106 M's', about 0.5 x 106 M's', about 1 x 105M-1s-1, about 0.5 x 105 M's', about 1 x 104 m-1S-1, about 0.5 x 104 M's', about 1 x 103 M's', or about 0.5 x 103 M-1s-1 (inclusive); about 0.5 x 103 M-1s-1 to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105M-1s-1, about 0.5 x 105 M's', about 1 x 104 M's', about 0.5 x 104 m-ls-1, or about 1 x 103 M-1s-1 (inclusive); about 1 x 103 M-1s-lto about 1 x 106 M-1s-1, about 0.5 x 106 M's', about 1 x 105M-1s-1, about 0.5 x 105 M's', about 1 x 104 M's', or about 0.5 x 104 m-ls-1 (inclusive); about 0.5 x 104 m-is-lto about 1 x 106M-1s-1, about 0.5 x 106 M's', about 1 x 105M-1s-1, about 0.5 x 105 M's', or about 1 x 104 M-1s-1 (inclusive);
about 1 x 104 M's' to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105M-is-i, or about 0.5 x 105 M's' (inclusive); about 0.5 x 105 M-1s-lto about 1 x 106M-1s-1, about 0.5 x 106 M's', or about 1 x 105 M's' (inclusive); about 1 x 105M-1s-1 to about 1 x 106 M's', or about 0.5 x 106 m-ls-1 (inclusive); or about 0.5 x 106 to about 1 X 106 M's' (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

Fusion Proteins In some embodiments, the IL-12/IL-23 inhibitor is a fusion protein, a soluble antagonist, or an antimicrobial peptide. In some embodiments, the fusion protein comprises a soluble fragment of a receptor of IL-12 or a soluble fragment of a receptor of IL-23. In some embodiments, the fusion protein comprises an extracellular domain of a receptor of IL-12 or an extracellular domain of a receptor of IL-23.
In some embodiments, the fusion protein is adnectin or a variant thereof (Tang et al., Immunology 135(2):112-124, 2012). In some embodiments, the soluble antagonist is a human IL-23Ra-chain mRNA transcript (Raymond et al., I Immunol. 185(12):7302-7308, .. 2010). In some embodiments, the IL-12/IL-23 is an antimicrobial peptide (e.g., MP-196 (Wenzel et al., PNAS 111(14):E1409-E1418, 2014)).
Small Molecules In some embodiments, the IL-12/IL-23 inhibitor is a small molecule. In some .. embodiments, the small molecule is STA-5326 (apilimod) or a variant thereof (Keino et al., Arthritis Res. Ther. 10: R122, 2008; Wada et al., Blood 109(3):1156-1164, 2007; Sands et al., Inflamm. Bowel Dis. 16(7):1209-1218, 2010).
TNFa Inhibitors The term "TNFa inhibitor" refers to an agent which directly or indirectly inhibits, impairs, reduces, down-regulates, or blocks TNFa activity and/or expression.
In some embodiments, a TNFa inhibitor is an inhibitory nucleic acid, an antibody or an antigen-binding fragment thereof, a fusion protein, a soluble TNFa receptor (a soluble TNFR1 or a soluble TNFR2), or a small molecule TNFa antagonist. In some embodiments, the inhibitory nucleic acid is a ribozyme, small hairpin RNA, a small interfering RNA, an antisense nucleic acid, or an aptamer.
Exemplary TNFa inhibitors that directly inhibit, impair, reduce, down-regulate, or block TNFa activity and/or expression can, e.g., inhibit or reduce binding of TNFa to its receptor (TNFR1 and/or TNFR2) and/or inhibit or decrease the expression level of TNFa or a receptor of TNFa (TNFR1 or TNFR2) in a cell (e.g., a mammalian cell). Non-limiting examples of TNFa inhibitors that directly inhibit, impair, reduce, down-regulate, or block TNFa activity and/or expression include inhibitory nucleic acids (e.g., any of the examples of inhibitory nucleic acids described herein), an antibody or fragment thereof, a fusion protein, a soluble TNFa receptor (e.g., a soluble TNFR1 or soluble TNFR2), and a small molecule TNFa antagonist.
Exemplary TNFa inhibitors that can indirectly inhibit, impair, reduce, down-regulate, or block TNFa activity and/or expression can, e.g., inhibit or decrease the level of downstream signaling of a TNFa receptor (e.g., TNFR1 or TNFR2) in a mammalian cell (e.g., decrease the level and/or activity of one or more of the following signaling proteins:
TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, and NF-KB in a mammalian cell), and/or decrease the level of TNFa-induced gene expression in a mammalian cell (e.g., decrease the transcription of genes regulated by, e.g., one or more transcription factors selected from the group of NF-KB, c-Jun, and ATF2). A
description of downstream signaling of a TNFa receptor is provided in Waj ant et al., Cell Death Differentiation 10:45-65, 2003 (incorporated herein by reference). For example, such indirect TNFa inhibitors can be an inhibitory nucleic acid that targets (decreases the expression) a signaling component downstream of a TNFa receptor (e.g., any one or more of the signaling components downstream of a TNFa receptor described herein or known in the art), a TNFa-induced gene (e.g., any TNFa-induced gene known in the art), or a transcription factor selected from the group of NF-KB, c-Jun, and ATF2.
In other examples, such indirect TNFa inhibitors can be a small molecule inhibitor of a signaling component downstream of a TNFa receptor (e.g., any of the signaling components downstream of a TNFa receptor described herein or known in the art), a small molecule inhibitor of a protein encoded by a TNFa-induced gene (e.g., any protein encoded by a TNFa-induced gene known in the art), and a small molecule inhibitor of a transcription factor selected from the group of NF-KB, c-Jun, and ATF2.
In other embodiments, TNFa inhibitors that can indirectly inhibit, impair, reduce, down-regulate, or block one or more components in a mammalian cell (e.g., a macrophage, a CD4+ lymphocyte, a NK cell, a neutrophil, a mast cell, a eosinophil, or a neuron) that are involved in the signaling pathway that results in TNFa mRNA transcription, TNFa mRNA
stabilization, and TNFa mRNA translation (e.g., one or more components selected from the group of CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, and MK2). For example, such indirect TNFa inhibitors can be an inhibitory nucleic acid that targets (decreases the expression) of a component in a mammalian cell that is involved in the signaling pathway that results in TNFa mRNA transcription, TNFa mRNA

stabilization, and TNFa mRNA translation (e.g., a component selected from the group of CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, INK, c-jun, MEK3/6, p38, PKR, TTP, and MK2). In other examples, an indirect TNFa inhibitors is a small molecule inhibitor of a component in a mammalian cell that is involved in the signaling pathway that results in TNFa mRNA transcription, TNFa mRNA stabilization, and TNFa mRNA translation (e.g., a component selected from the group of CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, 1KB, NF-KB, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, and MK2).
Inhibitory Nucleic Acids Inhibitory nucleic acids that can decrease the expression of TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA expression in a mammalian cell include antisense nucleic acid molecules, i.e., nucleic acid molecules whose nucleotide sequence is complementary to all or part of a TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, INK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA (e.g., complementary to all or a part of any one of SEQ ID NOs: 13-49).
Human TNFa CDS (SEQ ID NO: 13) ATGAGCACTGAAAGCATGATCCGGGACGTGGAGCTGGCCGAGGAGGCGCTCCCCAAGAAGACAGG
GGGGCCCCAGGGCTCCAGGCGGTGCTTGTTCCTCAGCCTCTTCTCCTTCCTGATCGTGGCAGGCGCC
ACCACGCTCTTCTGCCTGCTGCACTTTGGAGTGATCGGCCCCCAGAGGGAAGAGTTCCCCAGGGAC
CTCTCTCTAATCAGCCCTCTGGCCCAGGCAGTCAGATCATCTTCTCGAACCCCGAGTGACAAGCCTG
TAGCCCATGTTGTAGCAAACCCTCAAGCTGAGGGGCAGCTCCAGTGGCTGAACCGCCGGGCCAATG
CCCTCCTGGCCAATGGCGTGGAGCTGAGAGATAACCAGCTGGTGGTGCCATCAGAGGGCCTGTACC
TCATCTACTCCCAGGTCCTCTTCAAGGGCCAAGGCTGCCCCTCCACCCATGTGCTCCTCACCCACAC
CATCAGCCGCATCGCCGTCTCCTACCAGACCAAGGTCAACCTCCTCTCTGCCATCAAGAGCCCCTGC
CAGAGGGAGACCCCAGAGGGGGCTGAGGCCAAGCCCTGGTATGAGCCCATCTATCTGGGAGGGGT
CTTCCAGCTGGAGAAGGGTGACCGACTCAGCGCTGAGATCAATCGGCCCGACTATCTCGACTTTGC
CGAGTCTGGGCAGGTCTACTTTGGGATCATTGCCCTGTGA

Human TNFR1 CDS (SEQ ID NO: 14) ATGGGCCTCTCCACCGTGCCTGACCTGCTGCTGCCGCTGGTGCTCCTGGAGCTGTTGGTGGGAATAT
ACCCCTCAGGGGTTATTGGACTGGTCCCTCACCTAGGGGACAGGGAGAAGAGAGATAGTGTGTGTC
CCCAAGGAAAATATATCCACCCTCAAAATAATTCGATTTGCTGTACCAAGTGCCACAAAGGAACCTA
CTTGTACAATGACTGTCCAGGCCCGGGGCAGGATACGGACTGCAGGGAGTGTGAGAGCGGCTCCTT
CACCGCTTCAGAAAACCACCTCAGACACTGCCTCAGCTGCTCCAAATGCCGAAAGGAAATGGGTCA
GGTGGAGATCTCTTCTTGCACAGTGGACCGGGACACCGTGTGTGGCTGCAGGAAGAACCAGTACC
GGCATTATTGGAGTGAAAACCTTTTCCAGTGCTTCAATTGCAGCCTCTGCCTCAATGGGACCGTGCA
CCTCTCCTGCCAGGAGAAACAGAACACCGTGTGCACCTGCCATGCAGGTTTCTTTCTAAGAGAAAA
CGAGTGTGTCTCCTGTAGTAACTGTAAGAAAAGCCTGGAGTGCACGAAGTTGTGCCTACCCCAGAT
TGAGAATGTTAAGGGCACTGAGGACTCAGGCACCACAGTGCTGTTGCCCCTGGTCATTTTCTTTGGT
CTTTGCCTTTTATCCCTCCTCTTCATTGGTTTAATGTATCGCTACCAACGGTGGAAGTCCAAGCTCTAC
TCCATTGTTTGTGGGAAATCGACACCTGAAAAAGAGGGGGAGCTTGAAGGAACTACTACTAAGCCC
CTGGCCCCAAACCCAAGCTTCAGTCCCACTCCAGGCTTCACCCCCACCCTGGGCTTCAGTCCCGTG
CCCAGTTCCACCTTCACCTCCAGCTCCACCTATACCCCCGGTGACTGTCCCAACTTTGCGGCTCCCC
GCAGAGAGGTGGCACCACCCTATCAGGGGGCTGACCCCATCCTTGCGACAGCCCTCGCCTCCGACC
CCATCCCCAACCCCCTTCAGAAGTGGGAGGACAGCGCCCACAAGCCACAGAGCCTAGACACTGAT
GACCCCGCGACGCTGTACGCCGTGGTGGAGAACGTGCCCCCGTTGCGCTGGAAGGAATTCGTGCG
GCGCCTAGGGCTGAGCGACCACGAGATCGATCGGCTGGAGCTGCAGAACGGGCGCTGCCTGCGCG
AGGCGCAATACAGCATGCTGGCGACCTGGAGGCGGCGCACGCCGCGGCGCGAGGCCACGCTGGAG
CTGCTGGGACGCGTGCTCCGCGACATGGACCTGCTGGGCTGCCTGGAGGACATCGAGGAGGCGCTT
TGCGGCCCCGCCGCCCTCCCGCCCGCGCCCAGTCTTCTCAGATGA
Human TNFR2 CDS (SEQ ID NO: 15) ATGGCGCCCGTCGCCGTCTGGGCCGCGCTGGCCGTCGGACTGGAGCTCTGGGCTGCGGCGCACGCC
TTGCCCGCCCAGGTGGCATTTACACCCTACGCCCCGGAGCCCGGGAGCACATGCCGGCTCAGAGAA
TACTATGACCAGACAGCTCAGATGTGCTGCAGCAAATGCTCGCCGGGCCAACATGCAAAAGTCTTC
TGTACCAAGACCTCGGACACCGTGTGTGACTCCTGTGAGGACAGCACATACACCCAGCTCTGGAAC
TGGGTTCCCGAGTGCTTGAGCTGTGGCTCCCGCTGTAGCTCTGACCAGGTGGAAACTCAAGCCTGC
ACTCGGGAACAGAACCGCATCTGCACCTGCAGGCCCGGCTGGTACTGCGCGCTGAGCAAGCAGGA
GGGGTGCCGGCTGTGCGCGCCGCTGCGCAAGTGCCGCCCGGGCTTCGGCGTGGCCAGACCAGGAA
CTGAAACATCAGACGTGGTGTGCAAGCCCTGTGCCCCGGGGACGTTCTCCAACACGACTTCATCCA
CGGATATTTGCAGGCCCCACCAGATCTGTAACGTGGTGGCCATCCCTGGGAATGCAAGCATGGATGC
AGTCTGCACGTCCACGTCCCCCACCCGGAGTATGGCCCCAGGGGCAGTACACTTACCCCAGCCAGT
GTCCACACGATCCCAACACACGCAGCCAACTCCAGAACCCAGCACTGCTCCAAGCACCTCCTTCCT
GCTCCCAATGGGCCCCAGCCCCCCAGCTGAAGGGAGCACTGGCGACTTCGCTCTTCCAGTTGGACT
GATTGTGGGTGTGACAGCCTTGGGTCTACTAATAATAGGAGTGGTGAACTGTGTCATCATGACCCAG
GTGAAAAAGAAGCCCTTGTGCCTGCAGAGAGAAGCCAAGGTGCCTCACTTGCCTGCCGATAAGGC
CCGGGGTACACAGGGCCCCGAGCAGCAGCACCTGCTGATCACAGCGCCGAGCTCCAGCAGCAGCT
CCCTGGAGAGCTCGGCCAGTGCGTTGGACAGAAGGGCGCCCACTCGGAACCAGCCACAGGCACCA
GGCGTGGAGGCCAGTGGGGCCGGGGAGGCCCGGGCCAGCACCGGGAGCTCAGATTCTTCCCCTGG
TGGCCATGGGACCCAGGTCAATGTCACCTGCATCGTGAACGTCTGTAGCAGCTCTGACCACAGCTC
ACAGTGCTCCTCCCAAGCCAGCTCCACAATGGGAGACACAGATTCCAGCCCCTCGGAGTCCCCGAA
GGACGAGCAGGTCCCCTTCTCCAAGGAGGAATGTGCCTTTCGGTCACAGCTGGAGACGCCAGAGA
CCCTGCTGGGGAGCACCGAAGAGAAGCCCCTGCCCCTTGGAGTGCCTGATGCTGGGATGAAGCCC
AGTTAA
Human TRADD CDS (SEQ ID NO: 16) ATGGCAGCTGGGCAAAATGGGCACGAAGAGTGGGTGGGCAGCGCATACCTGTTTGTGGAGTCCTCG
CTGGACAAGGTGGTCCTGTCGGATGCCTACGCGCACCCCCAGCAGAAGGTGGCAGTGTACAGGGCT
CTGCAGGCTGCCTTGGCAGAGAGCGGCGGGAGCCCGGACGTGCTGCAGATGCTGAAGATCCACCG
CAGCGACCCGCAGCTGATCGTGCAGCTGCGATTCTGCGGGCGGCAGCCCTGTGGCCGCTTCCTCCG
CGCCTACCGCGAGGGGGCGCTGCGCGCCGCGCTGCAGAGGAGCCTGGCGGCCGCGCTCGCCCAGC
ACTCGGTGCCGCTGCAACTGGAGCTGCGCGCCGGCGCCGAGCGGCTGGACGCTTTGCTGGCGGAC
GAGGAGCGCTGTTTGAGTTGCATCCTAGCCCAGCAGCCCGACCGGCTCCGGGATGAAGAACTGGCT

GAGCTGGAGGATGCGCTGCGAAATCTGAAGTGCGGCTCGGGGGCCCGGGGTGGCGACGGGGAGGT
CGCTTCGGCCCCCTTGCAGCCCCCGGTGCCCTCTCTGTCGGAGGTGAAGCCGCCGCCGCCGCCGCC
ACCTGCCCAGACTTTTCTGTTCCAGGGTCAGCCTGTAGTGAATCGGCCGCTGAGCCTGAAGGACCA
ACAGACGTTCGCGCGCTCTGTGGGTCTCAAATGGCGCAAGGTGGGGCGCTCACTGCAGCGAGGCT
GCCGGGCGCTGCGGGACCCGGCGCTGGACTCGCTGGCCTACGAGTACGAGCGCGAGGGACTGTAC
GAGCAGGCCTTCCAGCTGCTGCGGCGCTTCGTGCAGGCCGAGGGCCGCCGCGCCACGCTGCAGCG
CCTGGTGGAGGCACTCGAGGAGAACGAGCTCACCAGCCTGGCAGAGGACTTGCTGGGCCTGACCG
ATCCCAATGGCGGCCTGGCCTAG
Human TRAF2 CDS (SEQ ID NO: 17) ATGGCTGCAGCTAGCGTGACCCCCCCTGGCTCCCTGGAGTTGCTACAGCCCGGCTTCTCCAAGACC
CTCCTGGGGACCAAGCTGGAAGCCAAGTACCTGTGCTCCGCCTGCAGAAACGTCCTCCGCAGGCCC
TTCCAGGCGCAGTGTGGCCACCGGTACTGCTCCTTCTGCCTGGCCAGCATCCTCAGCTCTGGGCCTC
AGAACTGTGCTGCCTGTGTTCACGAGGGCATATATGAAGAAGGCATTTCTATTTTAGAAAGCAGTTC
GGCCTTCCCAGATAATGCTGCCCGCAGGGAGGTGGAGAGCCTGCCGGCCGTCTGTCCCAGTGATGG
ATGCACCTGGAAGGGGACCCTGAAAGAATACGAGAGCTGCCACGAAGGCCGCTGCCCGCTCATGC
TGACCGAATGTCCCGCGTGCAAAGGCCTGGTCCGCCTTGGTGAAAAGGAGCGCCACCTGGAGCAC
GAGTGCCCGGAGAGAAGCCTGAGCTGCCGGCATTGCCGGGCACCCTGCTGCGGAGCAGACGTGAA
GGCGCACCACGAGGTCTGCCCCAAGTTCCCCTTAACTTGTGACGGCTGCGGCAAGAAGAAGATCCC
CCGGGAGAAGTTTCAGGACCACGTCAAGACTTGTGGCAAGTGTCGAGTCCCTTGCAGATTCCACGC
CATCGGCTGCCTCGAGACGGTAGAGGGTGAGAAACAGCAGGAGCACGAGGTGCAGTGGCTGCGGG
AGCACCTGGCCATGCTACTGAGCTCGGTGCTGGAGGCAAAGCCCCTCTTGGGAGACCAGAGCCAC
GCGGGGTCAGAGCTCCTGCAGAGGTGCGAGAGCCTGGAGAAGAAGACGGCCACTTTTGAGAACAT
TGTCTGCGTCCTGAACCGGGAGGTGGAGAGGGTGGCCATGACTGCCGAGGCCTGCAGCCGGCAGC
ACCGGCTGGACCAAGACAAGATTGAAGCCCTGAGTAGCAAGGTGCAGCAGCTGGAGAGGAGCATT
GGCCTCAAGGACCTGGCGATGGCTGACTTGGAGCAGAAGGTCTTGGAGATGGAGGCATCCACCTAC
GATGGGGTCTTCATCTGGAAGATCTCAGACTTCGCCAGGAAGCGCCAGGAAGCTGTGGCTGGCCGC
ATACCCGCCATCTTCTCCCCAGCCTTCTACACCAGCAGGTACGGCTACAAGATGTGTCTGCGTATCTA
CCTGAACGGCGACGGCACCGGGCGAGGAACACACCTGTCCCTCTTCTTTGTGGTGATGAAGGGCCC
GAATGACGCCCTGCTGCGGTGGCCCTTCAACCAGAAGGTGACCTTAATGCTGCTCGACCAGAATAA
CCGGGAGCACGTGATTGACGCCTTCAGGCCCGACGTGACTTCATCCTCTTTTCAGAGGCCAGTCAA
CGACATGAACATCGCAAGCGGCTGCCCCCTCTTCTGCCCCGTCTCCAAGATGGAGGCAAAGAATTC
CTACGTGCGGGACGATGCCATCTTCATCAAGGCCATTGTGGACCTGACAGGGCTCTAA
Human MEKK1 CDS (SEQ ID NO: 18) ATGGCGGCGGCGGCGGGGAATCGCGCCTCGTCGTCGGGATTCCCGGGCGCCAGGGCTACGAGCCCT
GAGGCAGGCGGCGGCGGAGGAGCCCTCAAGGCGAGCAGCGCGCCCGCGGCTGCCGCGGGACTGC
TGCGGGAGGCGGGCAGCGGGGGCCGCGAGCGGGCGGACTGGCGGCGGCGGCAGCTGCGCAAAGT
GCGGAGTGTGGAGCTGGACCAGCTGCCTGAGCAGCCGCTCTTCCTTGCCGCCTCACCGCCGGCCTC
CTCGACTTCCCCGTCGCCGGAGCCCGCGGACGCAGCGGGGAGTGGGACCGGCTTCCAGCCTGTGG
CGGTGCCGCCGCCCCACGGAGCCGCGAGCCGCGGCGGCGCCCACCTTACCGAGTCGGTGGCGGCG
CCGGACAGCGGCGCCTCGAGTCCCGCAGCGGCCGAGCCCGGGGAGAAGCGGGCGCCCGCCGCCG
AGCCGTCTCCTGCAGCGGCCCCCGCCGGTCGTGAGATGGAGAATAAAGAAACTCTCAAAGGGTTGC
ACAAGATGGATGATCGTCCAGAGGAACGAATGATCAGGGAGAAACTGAAGGCAACCTGTATGCCA
GCCTGGAAGCACGAATGGTTGGAAAGGAGAAATAGGCGAGGGCCTGTGGTGGTAAAACCAATCCC
AGTTAAAGGAGATGGATCTGAAATGAATCACTTAGCAGCTGAGTCTCCAGGAGAGGTCCAGGCAAG
TGCGGCTTCACCAGCTTCCAAAGGCCGACGCAGTCCTTCTCCTGGCAACTCCCCATCAGGTCGCAC
AGTGAAATCAGAATCTCCAGGAGTAAGGAGAAAAAGAGTTTCCCCAGTGCCTTTTCAGAGTGGCA
GAATCACACCACCCCGAAGAGCCCCTTCACCAGATGGCTTCTCACCATATAGCCCTGAGGAAACAA
ACCGCCGTGTTAACAAAGTGATGCGGGCCAGACTGTACTTACTGCAGCAGATAGGGCCTAACTCTTT
CCTGATTGGAGGAGACAGCCCAGACAATAAATACCGGGTGTTTATTGGGCCTCAGAACTGCAGCTG
TGCACGTGGAACATTCTGTATTCATCTGCTATTTGTGATGCTCCGGGTGTTTCAACTAGAACCTTCAG
ACCCAATGTTATGGAGAAAAACTTTAAAGAATTTTGAGGTTGAGAGTTTGTTCCAGAAATATCACAG
TAGGCGTAGCTCAAGGATCAAAGCTCCATCTCGTAACACCATCCAGAAGTTTGTTTCACGCATGTCA
AATTCTCATACATTGTCATCATCTAGTACTTCTACGTCTAGTTCAGAAAACAGCATAAAGGATGAAGA

GGAACAGATGTGTCCTATTTGCTTGTTGGGCATGCTTGATGAAGAAAGTCTTACAGTGTGTGAAGAC
GGCTGCAGGAACAAGCTGCACCACCACTGCATGTCAATTTGGGCAGAAGAGTGTAGAAGAAATAG
AGAACCTTTAATATGTCCCCTTTGTAGATCTAAGTGGAGATCTCATGATTTCTACAGCCACGAGTTGT
CAAGTCCTGTGGATTCCCCTTCTTCCCTCAGAGCTGCACAGCAGCAAACCGTACAGCAGCAGCCTT
TGGCTGGATCACGAAGGAATCAAGAGAGCAATTTTAACCTTACTCATTATGGAACTCAGCAAATCCC
TCCTGCTTACAAAGATTTAGCTGAGCCATGGATTCAGGTGTTTGGAATGGAACTCGTTGGCTGCTTAT
TTTCTAGAAACTGGAATGTGAGAGAGATGGCCCTCAGGCGTCTTTCCCATGATGTCAGTGGGGCCCT
GCTGTTGGCAAATGGGGAGAGCACTGGAAATTCTGGGGGCAGCAGTGGAAGCAGCCCGAGTGGGG
GAGCCACCAGTGGGTCTTCCCAGACCAGTATCTCAGGAGATGTGGTGGAGGCATGCTGCAGCGTTC
TGTCAATGGTCTGTGCTGACCCTGTCTACAAAGTGTACGTTGCTGCTTTAAAAACATTGAGAGCCAT
GCTGGTATATACTCCTTGCCACAGTTTAGCGGAAAGAATCAAACTTCAGAGACTTCTCCAGCCAGTT
GTAGACACCATCCTAGTCAAATGTGCAGATGCCAATAGCCGCACAAGTCAGCTGTCCATATCAACAC
TGTTGGAACTGTGCAAAGGCCAAGCAGGAGAGTTGGCAGTTGGCAGAGAAATACTAAAAGCTGGA
TCCATTGGTATTGGTGGTGTTGATTATGTCTTAAATTGTATTCTTGGAAACCAAACTGAATCAAACAA
TTGGCAAGAACTTCTTGGCCGCCTTTGTCTTATAGATAGACTGTTGTTGGAATTTCCTGCTGAATTTT
ATCCTCATATTGTCAGTACTGATGTTTCACAAGCTGAGCCTGTTGAAATCAGGTATAAGAAGCTGCTG
TCCCTCTTAACCTTTGCTTTGCAGTCCATTGATAATTCCCACTCAATGGTTGGCAAACTTTCCAGAAG
GATCTACTTGAGTTCTGCAAGAATGGTTACTACAGTACCCCATGTGTTTTCAAAACTGTTAGAAATGC
TGAGTGTTTCCAGTTCCACTCACTTCACCAGGATGCGTCGCCGTTTGATGGCTATTGCAGATGAGGT
GGAAATTGCCGAAGCCATCCAGTTGGGCGTAGAAGACACTTTGGATGGTCAACAGGACAGCTTCTT
GCAGGCATCTGTTCCCAACAACTATCTGGAAACCACAGAGAACAGTTCCCCTGAGTGCACAGTCCA
TTTAGAGAAAACTGGAAAAGGATTATGTGCTACAAAATTGAGTGCCAGTTCAGAGGACATTTCTGA
GAGACTGGCCAGCATTTCAGTAGGACCTTCTAGTTCAACAACAACAACAACAACAACAACAGAGC
AACCAAAGCCAATGGTTCAAACAAAAGGCAGACCCCACAGTCAGTGTTTGAACTCCTCTCCTTTAT
CTCATCATTCCCAATTAATGTTTCCAGCCTTGTCAACCCCTTCTTCTTCTACCCCATCTGTACCAGCTG
GCACTGCAACAGATGTCTCTAAGCATAGACTTCAGGGATTCATTCCCTGCAGAATACCTTCTGCATCT
CCTCAAACACAGCGCAAGTTTTCTCTACAATTCCACAGAAACTGTCCTGAAAACAAAGACTCAGAT
AAACTTTCCCCAGTCTTTACTCAGTCAAGACCCTTGCCCTCCAGTAACATACACAGGCCAAAGCCAT
CTAGACCTACCCCAGGTAATACAAGTAAACAGGGAGATCCCTCAAAAAATAGCATGACACTTGATCT
GAACAGTAGTTCCAAATGTGATGACAGCTTTGGCTGTAGCAGCAATAGTAGTAATGCTGTTATACCC
AGTGACGAGACAGTGTTCACCCCAGTAGAGGAGAAATGCAGATTAGATGTCAATACAGAGCTCAAC
TCCAGTATTGAGGACCTTCTTGAAGCATCTATGCCTTCAAGTGATACAACAGTAACTTTTAAGTCAG
AAGTTGCTGTCCTGTCTCCTGAAAAGGCTGAAAATGATGATACCTACAAAGATGATGTGAATCATAA
TCAAAAGTGCAAAGAGAAGATGGAAGCTGAAGAAGAAGAAGCTTTAGCAATTGCCATGGCAATGT
CAGCGTCTCAGGATGCCCTCCCCATAGTTCCTCAGCTGCAGGTTGAAAATGGAGAAGATATCATCAT
TATTCAACAGGATACACCAGAGACTCTACCAGGACATACCAAAGCAAAACAACCGTATAGAGAAGA
CACTGAATGGCTGAAAGGTCAACAGATAGGCCTTGGAGCATTTTCTTCTTGTTATCAGGCTCAAGAT
GTGGGAACTGGAACTTTAATGGCTGTTAAACAGGTGACTTATGTCAGAAACACATCTTCTGAGCAA
GAAGAAGTAGTAGAAGCACTAAGAGAAGAGATAAGAATGATGAGCCATCTGAATCATCCAAACATC
ATTAGGATGTTGGGAGCCACGTGTGAGAAGAGCAATTACAATCTCTTCATTGAATGGATGGCAGGGG
GATCGGTGGCTCATTTGCTGAGTAAATATGGAGCCTTCAAAGAATCAGTAGTTATTAACTACACTGAA
CAGTTACTCCGTGGCCTTTCGTATCTCCATGAAAACCAAATCATTCACAGAGATGTCAAAGGTGCCA
ATTTGCTAATTGACAGCACTGGTCAGAGACTAAGAATTGCAGATTTTGGAGCTGCAGCCAGGTTGGC
ATCAAAAGGAACTGGTGCAGGAGAGTTTCAGGGACAATTACTGGGGACAATTGCATTTATGGCACC
TGAGGTACTAAGAGGTCAACAGTATGGAAGGAGCTGTGATGTATGGAGTGTTGGCTGTGCTATTATA
GAAATGGCTTGTGCAAAACCACCATGGAATGCAGAAAAACACTCCAATCATCTTGCTTTGATATTTA
AGATTGCTAGTGCAACTACTGCTCCATCGATCCCTTCACATTTGTCTCCTGGTTTACGAGATGTGGCT
CTTCGTTGTTTAGAACTTCAACCTCAGGACAGACCTCCATCAAGAGAGCTACTGAAGCATCCAGTCT
TTCGTACTACATGGTAG
Human MEKK4 CDS (SEQ ID NO: 19) ATGAGAGAAGCCGCTGCCGCGCTGGTCCCTCCTCCCGCCTTTGCCGTCACGCCTGCCGCCGCCATG
GAGGAGCCGCCGCCACCGCCGCCGCCGCCACCACCGCCACCGGAACCCGAGACCGAGTCAGAACC
CGAGTGCTGCTTGGCGGCGAGGCAAGAGGGCACATTGGGAGATTCAGCTTGCAAGAGTCCTGAATC
TGATCTAGAAGACTTCTCCGATGAAACAAATACAGAGAATCTTTATGGTACCTCTCCCCCCAGCACA
CCTCGACAGATGAAACGCATGTCAACCAAACATCAGAGGAATAATGTGGGGAGGCCAGCCAGTCG
GTCTAATTTGAAAGAAAAAATGAATGCACCAAATCAGCCTCCACATAAAGACACTGGAAAAACAGT

GGAGAATGTGGAAGAATACAGCTATAAGCAGGAGAAAAAGATCCGAGCAGCTCTTAGAACAACAG
AGCGTGATCATAAAAAAAATGTACAGTGCTCATTCATGTTAGACTCAGTGGGTGGATCTTTGCCAAA
AAAATCAATTCCAGATGTGGATCTCAATAAGCCTTACCTCAGCCTTGGCTGTAGCAATGCTAAGCTTC
CAGTATCTGTGCCCATGCCTATAGCCAGACCTGCACGCCAGACTTCTAGGACTGACTGTCCAGCAGA
TCGTTTAAAGTTTTTTGAAACTTTACGACTTTTGCTAAAGCTTACCTCAGTCTCAAAGAAAAAAGAC
AGGGAGCAAAGAGGACAAGAAAATACGTCTGGTTTCTGGCTTAACCGATCTAACGAACTGATCTGG
TTAGAGCTACAAGCCTGGCATGCAGGACGGACAATTAACGACCAGGACTTCTTTTTATATACAGCCC
GTCAAGCCATCCCAGATATTATTAATGAAATCCTTACTTTCAAAGTCGACTATGGGAGCTTCGCCTTT
GTTAGAGATAGAGCTGGTTTTAATGGTACTTCAGTAGAAGGGCAGTGCAAAGCCACTCCTGGAACA
AAGATTGTAGGTTACTCAACACATCATGAGCATCTCCAACGCCAGAGGGTCTCATTTGAGCAGGTAA
AACGGATAATGGAGCTGCTAGAGTACATAGAAGCACTTTATCCATCATTGCAGGCTCTTCAGAAGGA
CTATGAAAAATATGCTGCAAAAGACTTCCAGGACAGGGTGCAGGCACTCTGTTTGTGGTTAAACATC
ACAAAAGACTTAAATCAGAAATTAAGGATTATGGGCACTGTTTTGGGCATCAAGAATTTATCAGACA
TTGGCTGGCCAGTGTTTGAAATCCCTTCCCCTCGACCATCCAAAGGTAATGAGCCGGAGTATGAGGG
TGATGACACAGAAGGAGAATTAAAGGAGTTGGAAAGTAGTACGGATGAGAGTGAAGAAGAACAAA
TCTCTGATCCTAGGGTACCGGAAATCAGACAGCCCATAGATAACAGCTTCGACATCCAGTCGCGGGA
CTGCATATCCAAGAAGCTTGAGAGGCTCGAATCTGAGGATGATTCTCTTGGCTGGGGAGCACCAGA
CTGGAGCACAGAAGCAGGCTTTAGTAGACATTGTCTGACTTCTATTTATAGACCATTTGTAGACAAA
GCACTGAAGCAGATGGGGTTAAGAAAGTTAATTTTAAGACTTCACAAGCTAATGGATGGTTCCTTGC
AAAGGGCACGTATAGCATTGGTAAAGAACGATCGTCCAGTGGAGTTTTCTGAATTTCCAGATCCCAT
GTGGGGTTCAGATTATGTGCAGTTGTCAAGGACACCACCTTCATCTGAGGAGAAATGCAGTGCTGT
GTCGTGGGAGGAGCTGAAGGCCATGGATTTACCTTCATTCGAACCTGCCTTCCTAGTTCTCTGCCGA
GTCCTTCTGAATGTCATACATGAGTGTCTGAAGTTAAGATTGGAGCAGAGACCTGCTGGAGAACCAT
CTCTCTTGAGTATTAAGCAGCTGGTGAGAGAGTGTAAGGAGGTCCTGAAGGGCGGCCTGCTGATGA
AGCAGTACTACCAGTTCATGCTGCAGGAGGTTCTGGAGGACTTGGAGAAGCCCGACTGCAACATTG
ACGCTTTTGAAGAGGATCTACATAAAATGCTTATGGTGTATTTTGATTACATGAGAAGCTGGATCCAA
ATGCTACAGCAATTACCTCAAGCATCGCATAGTTTAAAAAATCTGTTAGAAGAAGAATGGAATTTCA
CCAAAGAAATAACTCATTACATACGGGGAGGAGAAGCACAGGCCGGGAAGCTTTTCTGTGACATTG
CAGGAATGCTGCTGAAATCTACAGGAAGTTTTTTAGAATTTGGCTTACAGGAGAGCTGTGCTGAATT
TTGGACTAGTGCGGATGACAGCAGTGCTTCCGACGAAATCAGGAGGTCTGTTATAGAGATCAGTCG
AGCCCTGAAGGAGCTCTTCCATGAAGCCAGAGAAAGGGCTTCCAAAGCACTTGGATTTGCTAAAAT
GTTGAGAAAGGACCTGGAAATAGCAGCAGAATTCAGGCTTTCAGCCCCAGTTAGAGACCTCCTGGA
TGTTCTGAAATCAAAACAGTATGTCAAGGTGCAAATTCCTGGGTTAGAAAACTTGCAAATGTTTGTT
CCAGACACTCTTGCTGAGGAGAAGAGTATTATTTTGCAGTTACTCAATGCAGCTGCAGGAAAGGAC
TGTTCAAAAGATTCAGATGACGTACTCATCGATGCCTATCTGCTTCTGACCAAGCACGGTGATCGAG
CCCGTGATTCAGAGGACAGCTGGGGCACCTGGGAGGCACAGCCTGTCAAAGTCGTGCCTCAGGTG
GAGACTGTTGACACCCTGAGAAGCATGCAGGTGGATAATCTTTTACTAGTTGTCATGCAGTCTGCGC
ATCTCACAATTCAGAGAAAAGCTTTCCAGCAGTCCATTGAGGGACTTATGACTCTGTGCCAGGAGC
AGACATCCAGTCAGCCGGTCATCGCCAAAGCTTTGCAGCAGCTGAAGAATGATGCATTGGAGCTAT
GCAACAGGATAAGCAATGCCATTGACCGCGTGGACCACATGTTCACATCAGAATTTGATGCTGAGGT
TGATGAATCTGAATCTGTCACCTTGCAACAGTACTACCGAGAAGCAATGATTCAGGGGTACAATTTT
GGATTTGAGTATCATAAAGAAGTTGTTCGTTTGATGTCTGGGGAGTTTAGACAGAAGATAGGAGACA
AATATATAAGCTTTGCCCGGAAGTGGATGAATTATGTCCTGACTAAATGTGAGAGTGGTAGAGGTAC
AAGACCCAGGTGGGCGACTCAAGGATTTGATTTTCTACAAGCAATTGAACCTGCCTTTATTTCAGCT
TTACCAGAAGATGACTTCTTGAGTTTACAAGCCTTGATGAATGAATGCATTGGCCATGTCATAGGAA
AACCACACAGTCCTGTTACAGGTTTGTACCTTGCCATTCATCGGAACAGCCCCCGTCCTATGAAGGT
ACCTCGATGCCATAGTGACCCTCCTAACCCACACCTCATTATCCCCACTCCAGAGGGATTCAGCACT
CGGAGCATGCCTTCCGACGCGCGGAGCCATGGCAGCCCTGCTGCTGCTGCTGCTGCTGCTGCTGCT
GCTGTTGCTGCCAGTCGGCCCAGCCCCTCTGGTGGTGACTCTGTGCTGCCCAAATCCATCAGCAGTG
CCCATGATACCAGGGGTTCCAGCGTTCCTGAAAATGATCGATTGGCTTCCATAGCTGCTGAATTGCA
GTTTAGGTCCCTGAGTCGTCACTCAAGCCCCACGGAGGAGCGAGATGAACCAGCATATCCAAGAGG
AGATTCAAGTGGGTCCACAAGAAGAAGTTGGGAACTTCGGACACTAATCAGCCAGAGTAAAGATAC
TGCTTCTAAACTAGGACCCATAGAAGCTATCCAGAAGTCAGTCCGATTGTTTGAAGAAAAGAGGTA
CCGAGAAATGAGGAGAAAGAATATCATTGGTCAAGTTTGTGATACGCCTAAGTCCTATGATAATGTTA
TGCACGTTGGCTTGAGGAAGGTGACCTTCAAATGGCAAAGAGGAAACAAAATTGGAGAAGGCCAG
TATGGGAAGGTGTACACCTGCATCAGCGTCGACACCGGGGAGCTGATGGCCATGAAAGAGATTCGA
TTTCAACCTAATGACCATAAGACTATCAAGGAAACTGCAGACGAATTGAAAATATTCGAAGGCATCA
AACACCCCAATCTGGTTCGGTATTTTGGTGTGGAGCTCCATAGAGAAGAAATGTACATCTTCATGGA
GTACTGCGATGAGGGGACTTTAGAAGAGGTGTCAAGGCTGGGACTTCAGGAACATGTGATTAGGCT

GTATTCAAAGCAGATCACCATTGCGATCAACGTCCTCCATGAGCATGGCATAGTCCACCGTGACATT
AAAGGTGCCAATATCTTCCTTACCTCATCTGGATTAATCAAACTGGGAGATTTTGGATGTTCAGTAAA
GCTCAAAAACAATGCCCAGACCATGCCTGGTGAAGTGAACAGCACCCTGGGGACAGCAGCATACAT
GGCACCTGAAGTCATCACTCGTGCCAAAGGAGAGGGCCATGGGCGTGCGGCCGACATCTGGAGTCT
GGGGTGTGTTGTCATAGAGATGGTGACTGGCAAGAGGCCTTGGCATGAGTATGAGCACAACTTTCA
AATTATGTATAAAGTGGGGATGGGACATAAGCCACCAATCCCTGAAAGATTAAGCCCTGAAGGAAA
GGACTTCCTTTCTCACTGCCTTGAGAGTGACCCAAAGATGAGATGGACCGCCAGCCAGCTCCTCGA
CCATTCGTTTGTCAAGGTTTGCACAGATGAAGAATGA
Human MEKK7 CDS (SEQ ID NO: 20) ATGTCTACAGCCTCTGCCGCCTCCTCCTCCTCCTCGTCTTCGGCCGGTGAGATGATCGAAGCCCCTT
CCCAGGTCCTCAACTTTGAAGAGATCGACTACAAGGAGATCGAGGTGGAAGAGGTTGTTGGAAGA
GGAGCCTTTGGAGTTGTTTGCAAAGCTAAGTGGAGAGCAAAAGATGTTGCTATTAAACAAATAGAA
AGTGAATCTGAGAGGAAAGCGTTTATTGTAGAGCTTCGGCAGTTATCCCGTGTGAACCATCCTAATA
TTGTAAAGCTTTATGGAGCCTGCTTGAATCCAGTGTGTCTTGTGATGGAATATGCTGAAGGGGGCTC
TTTATATAATGTGCTGCATGGTGCTGAACCATTGCCATATTATACTGCTGCCCACGCAATGAGTTGGTG
TTTACAGTGTTCCCAAGGAGTGGCTTATCTTCACAGCATGCAACCCAAAGCGCTAATTCACAGGGAC
CTGAAACCACCAAACTTACTGCTGGTTGCAGGGGGGACAGTTCTAAAAATTTGTGATTTTGGTACA
GCCTGTGACATTCAGACACACATGACCAATAACAAGGGGAGTGCTGCTTGGATGGCACCTGAAGTT
TTTGAAGGTAGTAATTACAGTGAAAAATGTGACGTCTTCAGCTGGGGTATTATTCTTTGGGAAGTGA
TAACGCGTCGGAAACCCTTTGATGAGATTGGTGGCCCAGCTTTCCGAATCATGTGGGCTGTTCATAA
TGGTACTCGACCACCACTGATAAAAAATTTACCTAAGCCCATTGAGAGCCTGATGACTCGTTGTTGG
TCTAAAGATCCTTCCCAGCGCCCTTCAATGGAGGAAATTGTGAAAATAATGACTCACTTGATGCGGT
ACTTTCCAGGAGCAGATGAGCCATTACAGTATCCTTGTCAGTATTCAGATGAAGGACAGAGCAACTC
TGCCACCAGTACAGGCTCATTCATGGACATTGCTTCTACAAATACGAGTAACAAAAGTGACACTAAT
ATGGAGCAAGTTCCTGCCACAAATGATACTATTAAGCGCTTAGAATCAAAATTGTTGAAAAATCAGG
CAAAGCAACAGAGTGAATCTGGACGTTTAAGCTTGGGAGCCTCCCGTGGGAGCAGTGTGGAGAGC
TTGCCCCCAACCTCTGAGGGCAAGAGGATGAGTGCTGACATGTCTGAAATAGAAGCTAGGATCGCC
GCAACCACAGGCAACGGACAGCCAAGACGTAGATCCATCCAAGACTTGACTGTAACTGGAACAGA
ACCTGGTCAGGTGAGCAGTAGGTCATCCAGTCCCAGTGTCAGAATGATTACTACCTCAGGACCAAC
CTCAGAAAAGCCAACTCGAAGTCATCCATGGACCCCTGATGATTCCACAGATACCAATGGATCAGAT
AACTCCATCCCAATGGCTTATCTTACACTGGATCACCAACTACAGCCTCTAGCACCGTGCCCAAACT
CCAAAGAATCTATGGCAGTGTTTGAACAGCATTGTAAAATGGCACAAGAATATATGAAAGTTCAAAC
AGAAATTGCATTGTTATTACAGAGAAAGCAAGAACTAGTTGCAGAACTGGACCAGGATGAAAAGGA
CCAGCAAAATACATCTCGCCTGGTACAGGAACATAAAAAGCTTTTAGATGAAAACAAAAGCCTTTCT
ACTTACTACCAGCAATGCAAAAAACAACTAGAGGTCATCAGAAGTCAGCAGCAGAAACGACAAGG
CACTTCATGA
Human JNK CDS (SEQ ID NO: 21) ATGAGCAGAAGCAAGCGTGACAACAATTTTTATAGTGTAGAGATTGGAGATTCTACATTCACAGTCC
TGAAACGATATCAGAATTTAAAACCTATAGGCTCAGGAGCTCAAGGAATAGTATGCGCAGCTTATGA
TGCCATTCTTGAAAGAAATGTTGCAATCAAGAAGCTAAGCCGACCATTTCAGAATCAGACTCATGCC
AAGCGGGCCTACAGAGAGCTAGTTCTTATGAAATGTGTTAATCACAAAAATATAATTGGCCTTTTGA
ATGTTTTCACACCACAGAAATCCCTAGAAGAATTTCAAGATGTTTACATAGTCATGGAGCTCATGGAT
GCAAATCTTTGCCAAGTGATTCAGATGGAGCTAGATCATGAAAGAATGTCCTACCTTCTCTATCAGAT
GCTGTGTGGAATCAAGCACCTTCATTCTGCTGGAATTATTCATCGGGACTTAAAGCCCAGTAATATAG
TAGTAAAATCTGATTGCACTTTGAAGATTCTTGACTTCGGTCTGGCCAGGACTGCAGGAACGAGTTT
TATGATGACGCCTTATGTAGTGACTCGCTACTACAGAGCACCCGAGGTCATCCTTGGCATGGGCTAC
AAGGAAAACGTTGACATTTGGTCAGTTGGGTGCATCATGGGAGAAATGATCAAAGGTGGTGTTTTG
TTCCCAGGTACAGATCATATTGATCAGTGGAATAAAGTTATTGAACAGCTTGGAACACCATGTCCTG
AATTCATGAAGAAACTGCAACCAACAGTAAGGACTTACGTTGAAAACAGACCTAAATATGCTGGAT
ATAGCTTTGAGAAACTCTTCCCTGATGTCCTTTTCCCAGCTGACTCAGAACACAACAAACTTAAAGC
CAGTCAGGCAAGGGATTTGTTATCCAAAATGCTGGTAATAGATGCATCTAAAAGGATCTCTGTAGAT
GAAGCTCTCCAACACCCGTACATCAATGTCTGGTATGATCCTTCTGAAGCAGAAGCTCCACCACCAA
AGATCCCTGACAAGCAGTTAGATGAAAGGGAACACACAATAGAAGAGTGGAAAGAATTGATATATA
AGGAAGTTATGGACTTGGAGGAGAGAACCAAGAATGGAGTTATACGGGGGCAGCCCTCTCCTTTAG

GTGCAGCAGTGATCAATGGCTCTCAGCATCCATCATCATCGTCGTCTGTCAATGATGTGTCTTCAATG
TCAACAGATCCGACTTTGGCCTCTGATACAGACAGCAGTCTAGAAGCAGCAGCTGGGCCTCTGGGC
TGCTGTAGATGA
Human AP-1 CDS (SEQ ID NO: 22) ATGACTGCAAAGATGGAAACGACCTTCTATGACGATGCCCTCAACGCCTCGTTCCTCCCGTCCGAGA
GCGGACCTTATGGCTACAGTAACCCCAAGATCCTGAAACAGAGCATGACCCTGAACCTGGCCGACC
CAGTGGGGAGCCTGAAGCCGCACCTCCGCGCCAAGAACTCGGACCTCCTCACCTCGCCCGACGTG
GGGCTGCTCAAGCTGGCGTCGCCCGAGCTGGAGCGCCTGATAATCCAGTCCAGCAACGGGCACATC
ACCACCACGCCGACCCCCACCCAGTTCCTGTGCCCCAAGAACGTGACAGATGAGCAGGAGGGCTT
CGCCGAGGGCTTCGTGCGCGCCCTGGCCGAACTGCACAGCCAGAACACGCTGCCCAGCGTCACGT
CGGCGGCGCAGCCGGTCAACGGGGCAGGCATGGTGGCTCCCGCGGTAGCCTCGGTGGCAGGGGGC
AGCGGCAGCGGCGGCTTCAGCGCCAGCCTGCACAGCGAGCCGCCGGTCTACGCAAACCTCAGCAA
CTTCAACCCAGGCGCGCTGAGCAGCGGCGGCGGGGCGCCCTCCTACGGCGCGGCCGGCCTGGCCT
TTCCCGCGCAACCCCAGCAGCAGCAGCAGCCGCCGCACCACCTGCCCCAGCAGATGCCCGTGCAG
CACCCGCGGCTGCAGGCCCTGAAGGAGGAGCCTCAGACAGTGCCCGAGATGCCCGGCGAGACACC
GCCCCTGTCCCCCATCGACATGGAGTCCCAGGAGCGGATCAAGGCGGAGAGGAAGCGCATGAGGA
ACCGCATCGCTGCCTCCAAGTGCCGAAAAAGGAAGCTGGAGAGAATCGCCCGGCTGGAGGAAAAA
GTGAAAACCTTGAAAGCTCAGAACTCGGAGCTGGCGTCCACGGCCAACATGCTCAGGGAACAGGT
GGCACAGCTTAAACAGAAAGTCATGAACCACGTTAACAGTGGGTGCCAACTCATGCTAACGCAGCA
GTTGCAAACATTTTGA
Human ASK! CDS (SEQ ID NO: 23) ATGAGCACGGAGGCGGACGAGGGCATCACTTTCTCTGTGCCACCCTTCGCCCCCTCGGGCTTCTGC
ACCATCCCCGAGGGCGGCATCTGCAGGAGGGGAGGAGCGGCGGCGGTGGGCGAGGGCGAGGAGC
ACCAGCTGCCACCGCCGCCGCCGGGCAGCTTCTGGAACGTGGAGAGCGCCGCTGCCCCTGGCATC
GGTTGTCCGGCGGCCACCTCCTCGAGCAGTGCCACCCGAGGCCGGGGCAGCTCTGTTGGCGGGGG
CAGCCGACGGACCACGGTGGCATATGTGATCAACGAAGCGAGCCAAGGGCAACTGGTGGTGGCCG
AGAGCGAGGCCCTGCAGAGCTTGCGGGAGGCGTGCGAGACAGTGGGCGCCACCCTGGAAACCCTG
CATTTTGGGAAACTCGACTTTGGAGAAACCACCGTGCTGGACCGCTTTTACAATGCAGATATTGCGG
TGGTGGAGATGAGCGATGCCTTCCGGCAGCCGTCCTTGTTTTACCACCTTGGGGTGAGAGAAAGTT
TCAGCATGGCCAACAACATCATCCTCTACTGTGATACTAACTCGGACTCTCTGCAGTCACTGAAGGA
AATAATTTGCCAGAAGAATACTATGTGCACTGGGAACTACACCTTTGTTCCTTACATGATAACTCCAC
ATAACAAAGTCTACTGCTGTGACAGCAGCTTCATGAAGGGGTTGACAGAGCTCATGCAACCGAACT
TCGAGCTGCTTCTTGGACCCATCTGCTTACCTCTTGTGGATCGTTTTATTCAACTTTTGAAGGTGGCA
CAAGCAAGTTCTAGCCAGTACTTCCGGGAATCTATACTCAATGACATCAGGAAAGCTCGTAATTTATA
CACTGGTAAAGAATTGGCAGCTGAGTTGGCAAGAATTCGGCAGCGAGTAGATAATATCGAAGTCTT
GACAGCAGATATTGTCATAAATCTGTTACTTTCCTACAGAGATATCCAGGACTATGATTCTATTGTGAA
GCTGGTAGAGACTTTAGAAAAACTGCCAACCTTTGATTTGGCCTCCCATCACCATGTGAAGTTTCAT
TATGCATTTGCACTGAATAGGAGAAATCTCCCTGGTGACAGAGCAAAAGCTCTTGATATTATGATTCC
CATGGTGCAAAGCGAAGGACAAGTTGCTTCAGATATGTATTGCCTAGTTGGTCGAATCTACAAAGAT
ATGTTTTTGGACTCTAATTTCACGGACACTGAAAGCAGAGACCATGGAGCTTCTTGGTTCAAAAAG
GCATTTGAATCTGAGCCAACACTACAGTCAGGAATTAATTATGCGGTCCTCCTCCTGGCAGCTGGAC
ACCAGTTTGAATCTTCCTTTGAGCTCCGGAAAGTTGGGGTGAAGCTAAGTAGTCTTCTTGGTAAAAA
GGGAAACTTGGAAAAACTCCAGAGCTACTGGGAAGTTGGATTTTTTCTGGGGGCCAGCGTCCTAGC
CAATGACCACATGAGAGTCATTCAAGCATCTGAAAAGCTTTTTAAACTGAAGACACCAGCATGGTA
CCTCAAGTCTATTGTAGAGACAATTTTAATATATAAGCATTTTGTGAAACTGACCACAGAACAGCCTG
TGGCCAAGCAAGAACTTGTGGACTTTTGGATGGATTTCCTGGTCGAGGCCACAAAGACAGATGTTA
CTGTGGTTAGGTTTCCAGTATTAATATTAGAACCAACCAAAATCTATCAACCTTCTTATTTGTCTATCA
ACAATGAAGTTGAGGAAAAGACAATCTCTATTTGGCACGTGCTTCCTGATGACAAGAAAGGTATAC
ATGAGTGGAATTTTAGTGCCTCTTCTGTCAGGGGAGTGAGTATTTCTAAATTTGAAGAAAGATGCTG
CTTTCTTTATGTGCTTCACAATTCTGATGATTTCCAAATCTATTTCTGTACAGAACTTCATTGTAAAAA
GTTTTTTGAGATGGTGAACACCATTACCGAAGAGAAGGGGAGAAGCACAGAGGAAGGAGACTGTG
AAAGTGACTTGCTGGAGTATGACTATGAATATGATGAAAATGGTGACAGAGTCGTTTTAGGAAAAGG
CACTTATGGGATAGTCTACGCAGGTCGGGACTTGAGCAACCAAGTCAGAATTGCTATTAAGGAAATC
CCAGAGAGAGACAGCAGATACTCTCAGCCCCTGCATGAAGAAATAGCATTGCATAAACACCTGAAG

CACAAAAATATTGTCCAGTATCTGGGCTCTTTCAGTGAGAATGGTTTCATTAAAATCTTCATGGAGCA
GGTCCCTGGAGGAAGTCTTTCTGCTCTCCTTCGTTCCAAATGGGGTCCATTAAAAGACAATGAGCAA
ACAATTGGCTTTTATACAAAGCAAATACTGGAAGGATTAAAATATCTCCATGACAATCAGATAGTTCA
CCGGGACATAAAGGGTGACAATGTGTTGATTAATACCTACAGTGGTGTTCTCAAGATCTCTGACTTC
GGAACATCAAAGAGGCTTGCTGGCATAAACCCCTGTACTGAAACTTTTACTGGTACCCTCCAGTATA
TGGCACCAGAAATAATAGATAAAGGACCAAGAGGCTACGGAAAAGCAGCAGACATCTGGTCTCTGG
GCTGTACAATCATTGAAATGGCCACAGGAAAACCCCCATTTTATGAACTGGGAGAACCACAAGCAG
CTATGTTCAAGGTGGGAATGTTTAAAGTCCACCCTGAGATCCCAGAGTCCATGTCTGCAGAGGCCAA
GGCATTCATACTGAAATGTTTTGAACCAGATCCTGACAAGAGAGCCTGTGCTAACGACTTGCTTGTT
GATGAGTTTTTAAAAGTTTCAAGCAAAAAGAAAAAGACACAACCTAAGCTTTCAGCTCTTTCAGCT
GGATCAAATGAATATCTCAGGAGTATATCCTTGCCGGTACCTGTGCTGGTGGAGGACACCAGCAGCA
GCAGTGAGTACGGCTCAGTTTCACCCGACACGGAGTTGAAAGTGGACCCCTTCTCTTTCAAAACAA
GAGCCAAGTCCTGCGGAGAAAGAGATGTCAAGGGAATTCGGACACTCTTTTTGGGCATTCCAGATG
AGAATTTTGAAGATCACAGTGCTCCTCCTTCCCCTGAAGAAAAAGATTCTGGATTCTTCATGCTGAG
GAAGGACAGTGAGAGGCGAGCTACCCTTCACAGGATCCTGACGGAAGACCAAGACAAAATTGTGA
GAAACCTAATGGAATCTTTAGCTCAGGGGGCTGAAGAACCGAAACTAAAATGGGAACACATCACAA
CCCTCATTGCAAGCCTCAGAGAATTTGTGAGATCCACTGACCGAAAAATCATAGCCACCACACTGTC
AAAGCTGAAACTGGAGCTGGACTTCGACAGCCATGGCATTAGCCAAGTCCAGGTGGTACTCTTTGG
TTTTCAAGATGCTGTCAATAAAGTTCTTCGGAATCATAACATCAAGCCGCACTGGATGTTTGCCTTAG
ACAGTATCATTCGGAAGGCGGTACAGACAGCCATTACCATCCTGGTTCCAGAACTAAGGCCACATTT
CAGCCTTGCATCTGAGAGTGATACTGCTGATCAAGAAGACTTGGATGTAGAAGATGACCATGAGGA
ACAGCCTTCAAATCAAACTGTCCGAAGACCTCAGGCTGTCATTGAAGATGCTGTGGCTACCTCAGG
CGTGAGCACGCTCAGTTCTACTGTGTCTCATGATTCCCAGAGTGCTCACCGGTCACTGAATGTACAG
CTTGGAAGGATGAAAATAGAAACCAATAGATTACTGGAAGAATTGGTTCGGAAAGAGAAAGAATTA
CAAGCACTCCTTCATCGAGCTATTGAAGAAAAAGACCAAGAAATTAAACACCTGAAGCTTAAGTCC
CAACCCATAGAAATTCCTGAATTGCCTGTATTTCATCTAAATTCTTCTGGCACAAATACTGAAGATTC
TGAACTTACCGACTGGCTGAGAGTGAATGGAGCTGATGAAGACACTATAAGCCGGTTTTTGGCTGA
AGATTATACACTATTGGATGTTCTCTACTATGTTACACGTGATGACTTAAAATGCTTGAGACTAAGGG
GAGGGATGCTGTGCACACTGTGGAAGGCTATCATTGACTTTCGAAACAAACAGACTTGA
Human RIP CDS (SEQ ID NO: 24) ATGTGGAGCAAACTGAATAATGAAGAGCACAATGAGCTGAGGGAAGTGGACGGCACCGCTAAGAA
GAATGGCGGCACCCTCTACTACATGGCGCCCGAGCACCTGAATGACGTCAACGCAAAGCCCACAGA
GAAGTCGGATGTGTACAGCTTTGCTGTAGTACTCTGGGCGATATTTGCAAATAAGGAGCCATATGAA
AATGCTATCTGTGAGCAGCAGTTGATAATGTGCATAAAATCTGGGAACAGGCCAGATGTGGATGACA
TCACTGAGTACTGCCCAAGAGAAATTATCAGTCTCATGAAGCTCTGCTGGGAAGCGAATCCGGAAG
CTCGGCCGACATTTCCTGGCATTGAAGAAAAATTTAGGCCTTTTTATTTAAGTCAATTAGAAGAAAG
TGTAGAAGAGGACGTGAAGAGTTTAAAGAAAGAGTATTCAAACGAAAATGCAGTTGTGAAGAGAA
TGCAGTCTCTTCAACTTGATTGTGTGGCAGTACCTTCAAGCCGGTCAAATTCAGCCACAGAACAGC
CTGGTTCACTGCACAGTTCCCAGGGACTTGGGATGGGTCCTGTGGAGGAGTCCTGGTTTGCTCCTTC
CCTGGAGCACCCACAAGAAGAGAATGAGCCCAGCCTGCAGAGTAAACTCCAAGACGAAGCCAACT
ACCATCTTTATGGCAGCCGCATGGACAGGCAGACGAAACAGCAGCCCAGACAGAATGTGGCTTACA
ACAGAGAGGAGGAAAGGAGACGCAGGGTCTCCCATGACCCTTTTGCACAGCAAAGACCTTACGAG
AATTTTCAGAATACAGAGGGAAAAGGCACTGCTTATTCCAGTGCAGCCAGTCATGGTAATGCAGTGC
ACCAGCCCTCAGGGCTCACCAGCCAACCTCAAGTACTGTATCAGAACAATGGATTATATAGCTCACA
TGGCTTTGGAACAAGACCACTGGATCCAGGAACAGCAGGTCCCAGAGTTTGGTACAGGCCAATTCC
AAGTCATATGCCTAGTCTGCATAATATCCCAGTGCCTGAGACCAACTATCTAGGAAATACACCCACCA
TGCCATTCAGCTCCTTGCCACCAACAGATGAATCTATAAAATATACCATATACAATAGTACTGGCATTC
AGATTGGAGCCTACAATTATATGGAGATTGGTGGGACGAGTTCATCACTACTAGACAGCACAAATAC
GAACTTCAAAGAAGAGCCAGCTGCTAAGTACCAAGCTATCTTTGATAATACCACTAGTCTGACGGAT
AAACACCTGGACCCAATCAGGGAAAATCTGGGAAAGCACTGGAAAAACTGTGCCCGTAAACTGGG
CTTCACACAGTCTCAGATTGATGAAATTGACCATGACTATGAGCGAGATGGACTGAAAGAAAAGGT
TTACCAGATGCTCCAAAAGTGGGTGATGAGGGAAGGCATAAAGGGAGCCACGGTGGGGAAGCTGG
CCCAGGCGCTCCACCAGTGTTCCAGGATCGACCTTCTGAGCAGCTTGATTTACGTCAGCCAGAACTA
A

Human MEKK 3 CDS (SEQ ID NO: 25) ATGGACGAACAGGAGGCATTGAACTCAATCATGAACGATCTGGTGGCCCTCCAGATGAACCGACGT
CACCGGATGCCTGGATATGAGACCATGAAGAACAAAGACACAGGTCACTCAAATAGGCAGAAAAA
ACACAACAGCAGCAGCTCAGCCCTTCTGAACAGCCCCACAGTAACAACAAGCTCATGTGCAGGGG
CCAGTGAGAAAAAGAAATTTTTGAGTGACGTCAGAATCAAGTTCGAGCACAACGGGGAGAGGCGA
ATTATAGCGTTCAGCCGGCCTGTGAAATATGAAGATGTGGAGCACAAGGTGACAACAGTATTTGGAC
AACCTCTTGATCTACATTACATGAACAATGAGCTCTCCATCCTGCTGAAAAACCAAGATGATCTTGAT
AAAGCAATTGACATTTTAGATAGAAGCTCAAGCATGAAAAGCCTTAGGATATTGCTGTTGTCCCAGG
ACAGAAACCATAACAGTTCCTCTCCCCACTCTGGGGTGTCCAGACAGGTGCGGATCAAGGCTTCCC
AGTCCGCAGGGGATATAAATACTATCTACCAGCCCCCCGAGCCCAGAAGCAGGCACCTCTCTGTCAG
CTCCCAGAACCCTGGCCGAAGCTCACCTCCCCCTGGCTATGTTCCTGAGCGGCAGCAGCACATTGC
CCGGCAGGGGTCCTACACCAGCATCAACAGTGAGGGGGAGTTCATCCCAGAGACCAGCGAGCAGT
GCATGCTGGATCCCCTGAGCAGTGCAGAAAATTCCTTGTCTGGAAGCTGCCAATCCTTGGACAGGT
CAGCAGACAGCCCATCCTTCCGGAAATCACGAATGTCCCGTGCCCAGAGCTTCCCTGACAACAGAC
AGGAATACTCAGATCGGGAAACTCAGCTTTATGACAAAGGGGTCAAAGGTGGAACCTACCCCCGGC
GCTACCACGTGTCTGTGCACCACAAGGACTACAGTGATGGCAGAAGAACATTTCCCCGAATACGGC
GTCATCAAGGCAACTTGTTCACCCTGGTGCCCTCCAGCCGCTCCCTGAGCACAAATGGCGAGAACA
TGGGTCTGGCTGTGCAATACCTGGACCCCCGTGGGCGCCTGCGGAGTGCGGACAGCGAGAATGCCC
TCTCTGTGCAGGAGAGGAATGTGCCAACCAAGTCTCCCAGTGCCCCCATCAACTGGCGCCGGGGAA
AGCTCCTGGGCCAGGGTGCCTTCGGCAGGGTCTATTTGTGCTATGACGTGGACACGGGACGTGAAC
TTGCTTCCAAGCAGGTCCAATTTGATCCAGACAGTCCTGAGACAAGCAAGGAGGTGAGTGCTCTGG
AGTGCGAGATCCAGTTGCTAAAGAACTTGCAGCATGAGCGCATCGTGCAGTACTATGGCTGTCTGCG
GGACCGCGCTGAGAAGACCCTGACCATCTTCATGGAGTACATGCCAGGGGGCTCGGTGAAAGACC
AGTTGAAGGCTTACGGTGCTCTGACAGAGAGCGTGACCCGAAAGTACACGCGGCAGATCCTGGAG
GGCATGTCCTACCTGCACAGCAACATGATTGTTCACCGGGACATTAAGGGAGCCAACATCCTCCGAG
ACTCTGCTGGGAATGTAAAGCTGGGGGACTTTGGGGCCAGCAAACGCCTGCAGACGATCTGTATGT
CGGGGACGGGCATGCGCTCCGTCACTGGCACACCCTACTGGATGAGCCCTGAGGTGATCAGCGGCG
AGGGCTATGGAAGGAAAGCAGACGTGTGGAGCCTGGGCTGCACTGTGGTGGAGATGCTGACAGAG
AAACCACCGTGGGCAGAGTATGAAGCTATGGCCGCCATCTTCAAGATTGCCACCCAGCCCACCAAT
CCTCAGCTGCCCTCCCACATCTCTGAACATGGCCGGGACTTCCTGAGGCGCATTTTTGTGGAGGCTC
GCCAGAGACCTTCAGCTGAGGAGCTGCTCACACACCACTTTGCACAGCTCATGTACTGA
Human MEKK 6 CDS (SEQ ID NO: 26) ATGGCGGGGCCGTGTCCCCGGTCCGGGGCGGAGCGCGCCGGCAGCTGCTGGCAGGACCCGCTGGC
CGTGGCGCTGAGCCGGGGCCGGCAGCTCGCGGCGCCCCCGGGCCGGGGCTGCGCGCGGAGCCGGC
CGCTCAGCGTGGTCTACGTGCTGACCCGGGAGCCGCAGCCCGGGCTCGAGCCTCGGGAGGGAACC
GAGGCGGAGCCGCTGCCCCTGCGCTGCCTGCGCGAGGCTTGCGCGCAGGTCCCCCGGCCGCGGCC
GCCCCCGCAGCTGCGCAGCCTGCCCTTCGGGACGCTGGAGCTAGGCGACACCGCGGCTCTGGATGC
CTTCTACAACGCGGATGTGGTGGTGCTGGAGGTGAGCAGCTCGCTGGTACAGCCCTCCCTGTTCTAC
CACCTTGGTGTGCGTGAGAGCTTCAGCATGACCAACAATGTGCTCCTCTGCTCCCAGGCCGACCTC
CCTGACCTGCAGGCCCTGCGGGAGGATGTTTTCCAGAAGAACTCGGATTGCGTTGGCAGCTACACA
CTGATCCCCTATGTGGTGACGGCCACTGGTCGGGTGCTGTGTGGTGATGCAGGCCTTCTGCGGGGCC
TGGCTGATGGGCTGGTACAGGCTGGAGTGGGGACCGAGGCCCTGCTCACTCCCCTGGTGGGCCGGC
TTGCCCGCCTGCTGGAGGCCACACCCACAGACTCTTGTGGCTATTTCCGGGAGACCATTCGGCGGG
ACATCCGGCAGGCGCGGGAGCGGTTCAGTGGGCCACAGCTGCGGCAGGAGCTGGCTCGCCTGCAG
CGGAGACTGGACAGCGTGGAGCTGCTGAGCCCCGACATCATCATGAACTTGCTGCTCTCCTACCGC
GATGTGCAGGACTACTCGGCCATCATTGAGCTGGTGGAGACGCTGCAGGCCTTGCCCACCTGTGAT
GTGGCCGAGCAGCATAATGTCTGCTTCCACTACACTTTTGCCCTCAACCGGAGGAACAGGCCTGGG
GACCGGGCGAAGGCCCTGTCTGTGCTGCTGCCGCTGGTACAGCTTGAGGGCTCTGTGGCGCCCGAT
CTGTACTGCATGTGTGGCCGTATCTACAAGGACATGTTCTTCAGCTCGGGTTTCCAGGATGCTGGGC
ACCGGGAGCAGGCCTATCACTGGTATCGCAAGGCTTTTGACGTAGAGCCCAGCCTTCACTCAGGCA
TCAATGCAGCTGTGCTCCTCATTGCTGCCGGGCAGCACTTTGAGGATTCCAAAGAGCTCCGGCTAAT
AGGCATGAAGCTGGGCTGCCTGCTGGCCCGCAAAGGCTGCGTGGAGAAGATGCAGTATTACTGGGA
TGTGGGTTTCTACCTGGGAGCCCAGATCCTCGCCAATGACCCCACCCAGGTGGTGCTGGCTGCAGA
GCAGCTGTATAAGCTCAATGCCCCCATATGGTACCTGGTGTCCGTGATGGAGACCTTCCTGCTCTACC
AGCACTTCAGGCCCACGCCAGAGCCCCCTGGAGGGCCACCACGCCGTGCCCACTTCTGGCTCCACT

TCTTGCTACAGTCCTGCCAACCATTCAAGACAGCCTGTGCCCAGGGCGACCAGTGCTTGGTGCTGG
TCCTGGAGATGAACAAGGTGCTGCTGCCTGCAAAGCTCGAGGTTCGGGGTACTGACCCAGTAAGCA
CAGTGACCCTGAGCCTGCTGGAGCCTGAGACCCAGGACATTCCCTCCAGCTGGACCTTCCCAGTCG
CCTCCATATGCGGAGTCAGCGCCTCAAAGCGCGACGAGCGCTGCTGCTTCCTCTATGCACTCCCCCC
GGCTCAGGACGTCCAGCTGTGCTTCCCCAGCGTAGGGCACTGCCAGTGGTTCTGCGGCCTGATCCA
GGCCTGGGTGACGAACCCGGATTCCACGGCGCCCGCGGAGGAGGCGGAGGGCGCGGGGGAGATGT
TGGAGTTTGATTATGAGTACACGGAGACGGGCGAGCGGCTGGTGCTGGGCAAGGGCACGTATGGGG
TGGTGTACGCGGGCCGCGATCGCCACACGAGGGTGCGCATCGCCATCAAGGAGATCCCGGAGCGG
GACAGCAGGTTCTCTCAGCCCCTGCATGAAGAGATCGCTCTTCACAGACGCCTGCGCCACAAGAAC
ATAGTGCGCTATCTGGGCTCAGCTAGCCAGGGCGGCTACCTTAAGATCTTCATGGAGGAAGTGCCTG
GAGGCAGCCTGTCCTCCTTGCTGCGGTCGGTGTGGGGACCCCTGAAGGACAACGAGAGCACCATC
AGTTTCTACACCCGCCAGATCCTGCAGGGACTTGGCTACTTGCACGACAACCACATCGTGCACAGG
GACATAAAAGGGGACAATGTGCTGATCAACACCTTCAGTGGGCTGCTCAAGATTTCTGACTTCGGC
ACCTCCAAGCGGCTGGCAGGCATCACACCTTGCACTGAGACCTTCACAGGAACTCTGCAGTATATG
GCCCCAGAAATCATTGACCAGGGCCCACGCGGGTATGGGAAAGCAGCTGACATCTGGTCACTGGGC
TGCACTGTCATTGAGATGGCCACAGGTCGCCCCCCCTTCCACGAGCTCGGGAGCCCACAGGCTGCC
ATGTTTCAGGTGGGTATGTACAAGGTCCATCCGCCAATGCCCAGCTCTCTGTCGGCCGAGGCCCAAG
CCTTTCTCCTCCGAACTTTTGAGCCAGACCCCCGCCTCCGAGCCAGCGCCCAGACACTGCTGGGGG
ACCCCTTCCTGCAGCCTGGGAAAAGGAGCCGCAGCCCCAGCTCCCCACGACATGCTCCACGGCCCT
CAGATGCCCCTTCTGCCAGTCCCACTCCTTCAGCCAACTCAACCACCCAGTCTCAGACATTCCCGTG
CCCTCAGGCACCCTCTCAGCACCCACCCAGCCCCCCGAAGCGCTGCCTCAGTTATGGGGGCACCAG
CCAGCTCCGGGTGCCCGAGGAGCCTGCGGCCGAGGAGCCTGCGTCTCCGGAGGAGAGTTCGGGGC
TGAGCCTGCTGCACCAGGAGAGCAAGCGTCGGGCCATGCTGGCCGCAGTATTGGAGCAGGAGCTG
CCAGCGCTGGCGGAGAATCTGCACCAGGAGCAGAAGCAAGAGCAGGGGGCCCGTCTGGGCAGAA
ACCATGTGGAAGAGCTGCTGCGCTGCCTCGGGGCACACATCCACACTCCCAACCGCCGGCAGCTCG
CCCAGGAGCTGCGGGCGCTGCAAGGACGGCTGAGGGCCCAGGGCCTTGGGCCTGCGCTTCTGCAC
AGACCGCTGTTTGCCTTCCCGGATGCGGTGAAGCAGATCCTCCGCAAGCGCCAGATCCGTCCACAC
TGGATGTTCGTTCTGGACTCACTGCTCAGCCGTGCTGTGCGGGCAGCCCTGGGTGTGCTAGGACCG
GAGGTGGAGAAGGAGGCGGTCTCACCGAGGTCAGAGGAGCTGAGTAATGAAGGGGACTCCCAGC
AGAGCCCAGGCCAGCAGAGCCCGCTTCCGGTGGAGCCCGAGCAGGGCCCCGCTCCTCTGATGGTG
CAGCTGAGCCTCTTGAGGGCAGAGACTGATCGGCTGCGCGAAATCCTGGCGGGGAAGGAACGGGA
GTACCAGGCCCTGGTGCAGCGGGCTCTACAGCGGCTGAATGAGGAAGCCCGGACCTATGTCCTGGC
CCCAGAGCCTCCAACTGCTCTTTCAACGGACCAGGGCCTGGTGCAGTGGCTACAGGAACTGAATGT
GGATTCAGGCACCATCCAAATGCTGTTGAACCATAGCTTCACCCTCCACACTCTGCTCACCTATGCC
ACTCGAGATGACCTCATCTACACCCGCATCAGGGGAGGGATGGTATGCCGCATCTGGAGGGCCATCT
TGGCACAGCGAGCAGGATCCACACCAGTCACCTCTGGACCCTGA
Human NIK CDS (SEQ ID NO: 27) ATGGCAGTGATGGAAATGGCCTGCCCAGGTGCCCCTGGCTCAGCAGTGGGGCAGCAGAAGGAACT
CCCCAAAGCCAAGGAGAAGACGCCGCCACTGGGGAAGAAACAGAGCTCCGTCTACAAGCTTGAG
GCCGTGGAGAAGAGCCCTGTGTTCTGCGGAAAGTGGGAGATCCTGAATGACGTGATTACCAAGGGC
ACAGCCAAGGAAGGCTCCGAGGCAGGGCCAGCTGCCATCTCTATCATCGCCCAGGCTGAGTGTGAG
AATAGCCAAGAGTTCAGCCCCACCTTTTCAGAACGCATTTTCATCGCTGGGTCCAAACAGTACAGCC
AGTCCGAGAGTCTTGATCAGATCCCCAACAATGTGGCCCATGCTACAGAGGGCAAAATGGCCCGTG
TGTGTTGGAAGGGAAAGCGTCGCAGCAAAGCCCGGAAGAAACGGAAGAAGAAGAGCTCAAAGTC
CCTGGCTCATGCAGGAGTGGCCTTGGCCAAACCCCTCCCCAGGACCCCTGAGCAGGAGAGCTGCA
CCATCCCAGTGCAGGAGGATGAGTCTCCACTCGGCGCCCCATATGTTAGAAACACCCCGCAGTTCAC
CAAGCCTCTGAAGGAACCAGGCCTTGGGCAACTCTGTTTTAAGCAGCTTGGCGAGGGCCTACGGCC
GGCTCTGCCTCGATCAGAACTCCACAAACTGATCAGCCCCTTGCAATGTCTGAACCACGTGTGGAA
ACTGCACCACCCCCAGGACGGAGGCCCCCTGCCCCTGCCCACGCACCCCTTCCCCTATAGCAGACT
GCCTCATCCCTTCCCATTCCACCCTCTCCAGCCCTGGAAACCTCACCCTCTGGAGTCCTTCCTGGGC
AAACTGGCCTGTGTAGACAGCCAGAAACCCTTGCCTGACCCACACCTGAGCAAACTGGCCTGTGTA
GACAGTCCAAAGCCCCTGCCTGGCCCACACCTGGAGCCCAGCTGCCTGTCTCGTGGTGCCCATGAG
AAGTTTTCTGTGGAGGAATACCTAGTGCATGCTCTGCAAGGCAGCGTGAGCTCAGGCCAGGCCCAC
AGCCTGACCAGCCTGGCCAAGACCTGGGCAGCAAGGGGCTCCAGATCCCGGGAGCCCAGCCCCAA
AACTGAGGACAACGAGGGTGTCCTGCTCACTGAGAAACTCAAGCCAGTGGATTATGAGTACCGAG
AAGAAGTCCACTGGGCCACGCACCAGCTCCGCCTGGGCAGAGGCTCCTTCGGAGAGGTGCACAGG

ATGGAGGACAAGCAGACTGGCTTCCAGTGCGCTGTCAAAAAGGTGCGGCTGGAAGTATTTCGGGC
AGAGGAGCTGATGGCATGTGCAGGATTGACCTCACCCAGAATTGTCCCTTTGTATGGAGCTGTGAG
AGAAGGGCCTTGGGTCAACATCTTCATGGAGCTGCTGGAAGGTGGCTCCCTGGGCCAGCTGGTCAA
GGAGCAGGGCTGTCTCCCAGAGGACCGGGCCCTGTACTACCTGGGCCAGGCCCTGGAGGGTCTGG
AATACCTCCACTCACGAAGGATTCTGCATGGGGACGTCAAAGCTGACAACGTGCTCCTGTCCAGCG
ATGGGAGCCACGCAGCCCTCTGTGACTTTGGCCATGCTGTGTGTCTTCAACCTGATGGCCTGGGAA
AGTCCTTGCTCACAGGGGACTACATCCCTGGCACAGAGACCCACATGGCTCCGGAGGTGGTGCTGG
GCAGGAGCTGCGACGCCAAGGTGGATGTCTGGAGCAGCTGCTGTATGATGCTGCACATGCTCAACG
GCTGCCACCCCTGGACTCAGTTCTTCCGAGGGCCGCTCTGCCTCAAGATTGCCAGCGAGCCTCCGC
CTGTGAGGGAGATCCCACCCTCCTGCGCCCCTCTCACAGCCCAGGCCATCCAAGAGGGGCTGAGGA
AAGAGCCCATCCACCGCGTGTCTGCAGCGGAGCTGGGAGGGAAGGTGAACCGGGCACTACAGCAA
GTGGGAGGTCTGAAGAGCCCTTGGAGGGGAGAATATAAAGAACCAAGACATCCACCGCCAAATCA
AGCCAATTACCACCAGACCCTCCATGCCCAGCCGAGAGAGCTTTCGCCAAGGGCCCCAGGGCCCCG
GCCAGCTGAGGAGACAACAGGCAGAGCCCCTAAGCTCCAGCCTCCTCTCCCACCAGAGCCCCCAG
AGCCAAACAAGTCTCCTCCCTTGACTTTGAGCAAGGAGGAGTCTGGGATGTGGGAACCCTTACCTC
TGTCCTCCCTGGAGCCAGCCCCTGCCAGAAACCCCAGCTCACCAGAGCGGAAAGCAACCGTCCCG
GAGCAGGAACTGCAGCAGCTGGAAATAGAATTATTCCTCAACAGCCTGTCCCAGCCATTTTCTCTGG
AGGAGCAGGAGCAAATTCTCTCGTGCCTCAGCATCGACAGCCTCTCCCTGTCGGATGACAGTGAGA
AGAACCCATCAAAGGCCTCTCAAAGCTCGCGGGACACCCTGAGCTCAGGCGTACACTCCTGGAGC
AGCCAGGCCGAGGCTCGAAGCTCCAGCTGGAACATGGTGCTGGCCCGGGGGCGGCCCACCGACAC
CCCAAGCTATTTCAATGGTGTGAAAGTCCAAATACAGTCTCTTAATGGTGAACACCTGCACATCCGG
GAGTTCCACCGGGTCAAAGTGGGAGACATCGCCACTGGCATCAGCAGCCAGATCCCAGCTGCAGCC
TTCAGCTTGGTCACCAAAGACGGGCAGCCTGTTCGCTACGACATGGAGGTGCCAGACTCGGGCATC
GACCTGCAGTGCACACTGGCCCCTGATGGCAGCTTCGCCTGGAGCTGGAGGGTCAAGCATGGCCAG
CTGGAGAACAGGCCCTAA
Human IKK CDS (SEQ ID NO: 28) ATGTTTTCAGGGGGGTGTCATAGCCCCGGGTTTGGCCGCCCCAGCCCCGCCTTCCCCGCCCCGGGG
AGCCCGCCCCCTGCCCCGCGTCCCTGCCGACAGGAAACAGGTGAGCAGATTGCCATCAAGCAGTGC
CGGCAGGAGCTCAGCCCCCGGAACCGAGAGCGGTGGTGCCTGGAGATCCAGATCATGAGAAGGCT
GACCCACCCCAATGTGGTGGCTGCCCGAGATGTCCCTGAGGGGATGCAGAACTTGGCGCCCAATGA
CCTGCCCCTGCTGGCCATGGAGTACTGCCAAGGAGGAGATCTCCGGAAGTACCTGAACCAGTTTGA
GAACTGCTGTGGTCTGCGGGAAGGTGCCATCCTCACCTTGCTGAGTGACATTGCCTCTGCGCTTAGA
TACCTTCATGAAAACAGAATCATCCATCGGGATCTAAAGCCAGAAAACATCGTCCTGCAGCAAGGA
GAACAGAGGTTAATACACAAAATTATTGACCTAGGATATGCCAAGGAGCTGGATCAGGGCAGTCTTT
GCACATCATTCGTGGGGACCCTGCAGTACCTGGCCCCAGAGCTACTGGAGCAGCAGAAGTACACAG
TGACCGTCGACTACTGGAGCTTCGGCACCCTGGCCTTTGAGTGCATCACGGGCTTCCGGCCCTTCCT
CCCCAACTGGCAGCCCGTGCAGTGGCATTCAAAAGTGCGGCAGAAGAGTGAGGTGGACATTGTTG
TTAGCGAAGACTTGAATGGAACGGTGAAGTTTTCAAGCTCTTTACCCTACCCCAATAATCTTAACAG
TGTCCTGGCTGAGCGACTGGAGAAGTGGCTGCAACTGATGCTGATGTGGCACCCCCGACAGAGGG
GCACGGATCCCACGTATGGGCCCAATGGCTGCTTCAAGGCCCTGGATGACATCTTAAACTTAAAGCT
GGTTCATATCTTGAACATGGTCACGGGCACCATCCACACCTACCCTGTGACAGAGGATGAGAGTCTG
CAGAGCTTGAAGGCCAGAATCCAACAGGACACGGGCATCCCAGAGGAGGACCAGGAGCTGCTGCA
GGAAGCGGGCCTGGCGTTGATCCCCGATAAGCCTGCCACTCAGTGTATTTCAGACGGCAAGTTAAAT
GAGGGCCACACATTGGACATGGATCTTGTTTTTCTCTTTGACAACAGTAAAATCACCTATGAGACTC
AGATCTCCCCACGGCCCCAACCTGAAAGTGTCAGCTGTATCCTTCAAGAGCCCAAGAGGAATCTCG
CCTTCTTCCAGCTGAGGAAGGTGTGGGGCCAGGTCTGGCACAGCATCCAGACCCTGAAGGAAGATT
GCAACCGGCTGCAGCAGGGACAGCGAGCCGCCATGATGAATCTCCTCCGAAACAACAGCTGCCTCT
CCAAAATGAAGAATTCCATGGCTTCCATGTCTCAGCAGCTCAAGGCCAAGTTGGATTTCTTCAAAAC
CAGCATCCAGATTGACCTGGAGAAGTACAGCGAGCAAACCGAGTTTGGGATCACATCAGATAAACT
GCTGCTGGCCTGGAGGGAAATGGAGCAGGCTGTGGAGCTCTGTGGGCGGGAGAACGAAGTGAAAC
TCCTGGTAGAACGGATGATGGCTCTGCAGACCGACATTGTGGACTTACAGAGGAGCCCCATGGGCC
GGAAGCAGGGGGGAACGCTGGACGACCTAGAGGAGCAAGCAAGGGAGCTGTACAGGAGACTAAG
GGAAAAACCTCGAGACCAGCGAACTGAGGGTGACAGTCAGGAAATGGTACGGCTGCTGCTTCAGG
CAATTCAGAGCTTCGAGAAGAAAGTGCGAGTGATCTATACGCAGCTCAGTAAAACTGTGGTTTGCA
AGCAGAAGGCGCTGGAACTGTTGCCCAAGGTGGAAGAGGTGGTGAGCTTAATGAATGAGGATGAG
AAGACTGTTGTCCGGCTGCAGGAGAAGCGGCAGAAGGAGCTCTGGAATCTCCTGAAGATTGCTTGT

AGCAAGGTCCGTGGTCCTGTCAGTGGAAGCCCGGATAGCATGAATGCCTCTCGACTTAGCCAGCCT
GGGCAGCTGATGTCTCAGCCCTCCACGGCCTCCAACAGCTTACCTGAGCCAGCCAAGAAGAGTGA
AGAACTGGTGGCTGAAGCACATAACCTCTGCACCCTGCTAGAAAATGCCATACAGGACACTGTGAG
GGAACAAGACCAGAGTTTCACGGCCCTAGACTGGAGCTGGTTACAGACGGAAGAAGAAGAGCACA
GCTGCCTGGAGCAGGCCTCATGA
Human NF-1(13 CDS (SEQ ID NO: 29) ATGGCAGAAGATGATCCATATTTGGGAAGGCCTGAACAAATGTTTCATTTGGATCCTTCTTTGACTCA
TACAATATTTAATCCAGAAGTATTTCAACCACAGATGGCACTGCCAACAGATGGCCCATACCTTCAA
ATATTAGAGCAACCTAAACAGAGAGGATTTCGTTTCCGTTATGTATGTGAAGGCCCATCCCATGGTGG
ACTACCTGGTGCCTCTAGTGAAAAGAACAAGAAGTCTTACCCTCAGGTCAAAATCTGCAACTATGT
GGGACCAGCAAAGGTTATTGTTCAGTTGGTCACAAATGGAAAAAATATCCACCTGCATGCCCACAG
CCTGGTGGGAAAACACTGTGAGGATGGGATCTGCACTGTAACTGCTGGACCCAAGGACATGGTGGT
CGGCTTCGCAAACCTGGGTATACTTCATGTGACAAAGAAAAAAGTATTTGAAACACTGGAAGCACG
AATGACAGAGGCGTGTATAAGGGGCTATAATCCTGGACTCTTGGTGCACCCTGACCTTGCCTATTTG
CAAGCAGAAGGTGGAGGGGACCGGCAGCTGGGAGATCGGGAAAAAGAGCTAATCCGCCAAGCAG
CTCTGCAGCAGACCAAGGAGATGGACCTCAGCGTGGTGCGGCTCATGTTTACAGCTTTTCTTCCGG
ATAGCACTGGCAGCTTCACAAGGCGCCTGGAACCCGTGGTATCAGACGCCATCTATGACAGTAAAG
CCCCCAATGCATCCAACTTGAAAATTGTAAGAATGGACAGGACAGCTGGATGTGTGACTGGAGGGG
AGGAAATTTATCTTCTTTGTGACAAAGTTCAGAAAGATGACATCCAGATTCGATTTTATGAAGAGGA
AGAAAATGGTGGAGTCTGGGAAGGATTTGGAGATTTTTCCCCCACAGATGTTCATAGACAATTTGCC
ATTGTCTTCAAAACTCCAAAGTATAAAGATATTAATATTACAAAACCAGCCTCTGTGTTTGTCCAGCT
TCGGAGGAAATCTGACTTGGAAACTAGTGAACCAAAACCTTTCCTCTACTATCCTGAAATCAAAGAT
AAAGAAGAAGTGCAGAGGAAACGTCAGAAGCTCATGCCCAATTTTTCGGATAGTTTCGGCGGTGGT
AGTGGTGCTGGAGCTGGAGGCGGAGGCATGTTTGGTAGTGGCGGTGGAGGAGGGGGCACTGGAAG
TACAGGTCCAGGGTATAGCTTCCCACACTATGGATTTCCTACTTATGGTGGGATTACTTTCCATCCTGG
AACTACTAAATCTAATGCTGGGATGAAGCATGGAACCATGGACACTGAATCTAAAAAGGACCCTGA
AGGTTGTGACAAAAGTGATGACAAAAACACTGTAAACCTCTTTGGGAAAGTTATTGAAACCACAGA
GCAAGATCAGGAGCCCAGCGAGGCCACCGTTGGGAATGGTGAGGTCACTCTAACGTATGCAACAG
GAACAAAAGAAGAGAGTGCTGGAGTTCAGGATAACCTCTTTCTAGAGAAGGCTATGCAGCTTGCAA
AGAGGCATGCCAATGCCCTTTTCGACTACGCGGTGACAGGAGACGTGAAGATGCTGCTGGCCGTCC
AGCGCCATCTCACTGCTGTGCAGGATGAGAATGGGGACAGTGTCTTACACTTAGCAATCATCCACCT
TCATTCTCAACTTGTGAGGGATCTACTAGAAGTCACATCTGGTTTGATTTCTGATGACATTATCAACA
TGAGAAATGATCTGTACCAGACGCCCTTGCACTTGGCAGTGATCACTAAGCAGGAAGATGTGGTGG
AGGATTTGCTGAGGGCTGGGGCCGACCTGAGCCTTCTGGACCGCTTGGGTAACTCTGTTTTGCACC
TAGCTGCCAAAGAAGGACATGATAAAGTTCTCAGTATCTTACTCAAGCACAAAAAGGCAGCACTAC
TTCTTGACCACCCCAACGGGGACGGTCTGAATGCCATTCATCTAGCCATGATGAGCAATAGCCTGCC
ATGTTTGCTGCTGCTGGTGGCCGCTGGGGCTGACGTCAATGCTCAGGAGCAGAAGTCCGGGCGCAC
AGCACTGCACCTGGCTGTGGAGCACGACAACATCTCATTGGCAGGCTGCCTGCTCCTGGAGGGTGA
TGCCCATGTGGACAGTACTACCTACGATGGAACCACACCCCTGCATATAGCAGCTGGGAGAGGGTCC
ACCAGGCTGGCAGCTCTTCTCAAAGCAGCAGGAGCAGATCCCCTGGTGGAGAACTTTGAGCCTCTC
TATGACCTGGATGACTCTTGGGAAAATGCAGGAGAGGATGAAGGAGTTGTGCCTGGAACCACGCCT
CTAGATATGGCCACCAGCTGGCAGGTATTTGACATATTAAATGGGAAACCATATGAGCCAGAGTTTAC
ATCTGATGATTTACTAGCACAAGGAGACATGAAACAGCTGGCTGAAGATGTGAAGCTGCAGCTGTAT
AAGTTACTAGAAATTCCTGATCCAGACAAAAACTGGGCTACTCTGGCGCAGAAATTAGGTCTGGGG
ATACTTAATAATGCCTTCCGGCTGAGTCCTGCTCCTTCCAAAACACTTATGGACAACTATGAGGTCTC
TGGGGGTACAGTCAGAGAGCTGGTGGAGGCCCTGAGACAAATGGGCTACACCGAAGCAATTGAAG
TGATCCAGGCAGCCTCCAGCCCAGTGAAGACCACCTCTCAGGCCCACTCGCTGCCTCTCTCGCCTG
CCTCCACAAGGCAGCAAATAGACGAGCTCCGAGACAGTGACAGTGTCTGCGACAGCGGCGTGGAG
ACATCCTTCCGCAAACTCAGCTTTACCGAGTCTCTGACCAGTGGTGCCTCACTGCTAACTCTCAACA
AAATGCCCCATGATTATGGGCAGGAAGGACCTCTAGAAGGCAAAATTTAG
Human CD14 CDS (SEQ ID NO: 30) ATGGAGCGCGCGTCCTGCTTGTTGCTGCTGCTGCTGCCGCTGGTGCACGTCTCTGCGACCACGCCA
GAACCTTGTGAGCTGGACGATGAAGATTTCCGCTGCGTCTGCAACTTCTCCGAACCTCAGCCCGAC
TGGTCCGAAGCCTTCCAGTGTGTGTCTGCAGTAGAGGTGGAGATCCATGCCGGCGGTCTCAACCTA

GAGCCGTTTCTAAAGCGCGTCGATGCGGACGCCGACCCGCGGCAGTATGCTGACACGGTCAAGGCT
CTCCGCGTGCGGCGGCTCACAGTGGGAGCCGCACAGGTTCCTGCTCAGCTACTGGTAGGCGCCCTG
CGTGTGCTAGCGTACTCCCGCCTCAAGGAACTGACGCTCGAGGACCTAAAGATAACCGGCACCATG
CCTCCGCTGCCTCTGGAAGCCACAGGACTTGCACTTTCCAGCTTGCGCCTACGCAACGTGTCGTGG
GCGACAGGGCGTTCTTGGCTCGCCGAGCTGCAGCAGTGGCTCAAGCCAGGCCTCAAGGTACTGAG
CATTGCCCAAGCACACTCGCCTGCCTTTTCCTGCGAACAGGTTCGCGCCTTCCCGGCCCTTACCAGC
CTAGACCTGTCTGACAATCCTGGACTGGGCGAACGCGGACTGATGGCGGCTCTCTGTCCCCACAAG
TTCCCGGCCATCCAGAATCTAGCGCTGCGCAACACAGGAATGGAGACGCCCACAGGCGTGTGCGCC
GCACTGGCGGCGGCAGGTGTGCAGCCCCACAGCCTAGACCTCAGCCACAACTCGCTGCGCGCCAC
CGTAAACCCTAGCGCTCCGAGATGCATGTGGTCCAGCGCCCTGAACTCCCTCAATCTGTCGTTCGCT
GGGCTGGAACAGGTGCCTAAAGGACTGCCAGCCAAGCTCAGAGTGCTCGATCTCAGCTGCAACAG
ACTGAACAGGGCGCCGCAGCCTGACGAGCTGCCCGAGGTGGATAACCTGACACTGGACGGGAATC
CCTTCCTGGTCCCTGGAACTGCCCTCCCCCACGAGGGCTCAATGAACTCCGGCGTGGTCCCAGCCT
GTGCACGTTCGACCCTGTCGGTGGGGGTGTCGGGAACCCTGGTGCTGCTCCAAGGGGCCCGGGGC
TTTGCCTAA
Human MyD88 CDS (SEQ ID NO: 31) AT GC GAC C C GAC C GC GC T GAGGC T C C AGGAC C GC C C GC CAT GGC T GCAGGAGGT
CCC GGCGC GGGGTCTGCGGC CC CGGTC TC CTCCACATCCTCC CTTC CCC TGGCTGC
TCTCAACATGCGAGTGCGGCGCCGCCTGTCTCTGTTCTTGAACGTGCGGACACAG
GTGGC GGC C GAC T GGAC C GC GC TGGC GGAGGAGAT GGAC T TT GAGTAC TT GGAG
AT C C GGCAAC TGGAGACACAAGC GGAC C C CAC T GGCAGGC TGC T GGAC GC C T GG
CAGGGACGCCCTGGCGCCTCTGTAGGCCGACTGCTCGAGCTGCTTACCAAGCTGG
GC C GC GAC GAC GT GC T GC T GGAGC T GGGAC C CAGC ATT GGTGC C GC C GGATGGT
GGTGGTTGTCTCTGATGATTACCTGCAGAGCAAGGAATGTGACTTCCAGACCAAA
TTTGCACTCAGCCTCTCTCCAGGTGCCCATCAGAAGCGACTGA
Human IRAK CDS (SEQ ID NO: 32) ATGGCCGGGGGGCCGGGCCCGGGGGAGCCCGCAGCCCCCGGCGCCCAGCACTTCTTGTACGAGGT
GCCGCCCTGGGTCATGTGCCGCTTCTACAAAGTGATGGACGCCCTGGAGCCCGCCGACTGGTGCCA
GTTCGCCGCCCTGATCGTGCGCGACCAGACCGAGCTGCGGCTGTGCGAGCGCTCCGGGCAGCGCA
CGGCCAGCGTCCTGTGGCCCTGGATCAACCGCAACGCCCGTGTGGCCGACCTCGTGCACATCCTCA
CGCACCTGCAGCTGCTCCGTGCGCGGGACATCATCACAGCCTGGCACCCTCCCGCCCCGCTTCCGT
CCCCAGGCACCACTGCCCCGAGGCCCAGCAGCATCCCTGCACCCGCCGAGGCCGAGGCCTGGAGC
CCCCGGAAGTTGCCATCCTCAGCCTCCACCTTCCTCTCCCCAGCTTTTCCAGGCTCCCAGACCCATT
CAGGGCCTGAGCTCGGCCTGGTCCCAAGCCCTGCTTCCCTGTGGCCTCCACCGCCATCTCCAGCCC
CTTCTTCTACCAAGCCAGGCCCAGAGAGCTCAGTGTCCCTCCTGCAGGGAGCCCGCCCCTTTCCGT
TTTGCTGGCCCCTCTGTGAGATTTCCCGGGGCACCCACAACTTCTCGGAGGAGCTCAAGATCGGGG
AGGGTGGCTTTGGGTGCGTGTACCGGGCGGTGATGAGGAACACGGTGTATGCTGTGAAGAGGCTGA
AGGAGAACGCTGACCTGGAGTGGACTGCAGTGAAGCAGAGCTTCCTGACCGAGGTGGAGCAGCTG
TCCAGGTTTCGTCACCCAAACATTGTGGACTTTGCTGGCTACTGTGCTCAGAACGGCTTCTACTGCC
TGGTGTACGGCTTCCTGCCCAACGGCTCCCTGGAGGACCGTCTCCACTGCCAGACCCAGGCCTGCC
CACCTCTCTCCTGGCCTCAGCGACTGGACATCCTTCTGGGTACAGCCCGGGCAATTCAGTTTCTACA
TCAGGACAGCCCCAGCCTCATCCATGGAGACATCAAGAGTTCCAACGTCCTTCTGGATGAGAGGCT
GACACCCAAGCTGGGAGACTTTGGCCTGGCCCGGTTCAGCCGCTTTGCCGGGTCCAGCCCCAGCCA
GAGCAGCATGGTGGCCCGGACACAGACAGTGCGGGGCACCCTGGCCTACCTGCCCGAGGAGTACA
TCAAGACGGGAAGGCTGGCTGTGGACACGGACACCTTCAGCTTTGGGGTGGTAGTGCTAGAGACC
TTGGCTGGTCAGAGGGCTGTGAAGACGCACGGTGCCAGGACCAAGTATCTGAAAGACCTGGTGGA
AGAGGAGGCTGAGGAGGCTGGAGTGGCTTTGAGAAGCACCCAGAGCACACTGCAAGCAGGTCTG
GCTGCAGATGCCTGGGCTGCTCCCATCGCCATGCAGATCTACAAGAAGCACCTGGACCCCAGGCCC
GGGCCCTGCCCACCTGAGCTGGGCCTGGGCCTGGGCCAGCTGGCCTGCTGCTGCCTGCACCGCCGG
GCCAAAAGGAGGCCTCCTATGACCCAGGAGAACTCCTACGTGTCCAGCACTGGCAGAGCCCACAG
TGGGGCTGCTCCATGGCAGCCCCTGGCAGCGCCATCAGGAGCCAGTGCCCAGGCAGCAGAGCAGC
TGCAGAGAGGCCCCAACCAGCCCGTGGAGAGTGACGAGAGCCTAGGCGGCCTCTCTGCTGCCCTG

CGCTCCTGGCACTTGACTCCAAGCTGCCCTCTGGACCCAGCACCCCTCAGGGAGGCCGGCTGTCCT
CAGGGGGACACGGCAGGAGAATCGAGCTGGGGGAGTGGCCCAGGATCCCGGCCCACAGCCGTGG
AAGGACTGGCCCTTGGCAGCTCTGCATCATCGTCGTCAGAGCCACCGCAGATTATCATCAACCCTGC
CCGACAGAAGATGGTCCAGAAGCTGGCCCTGTACGAGGATGGGGCCCTGGACAGCCTGCAGCTGC
TGTCGTCCAGCTCCCTCCCAGGCTTGGGCCTGGAACAGGACAGGCAGGGGCCCGAAGAAAGTGAT
GAATTTCAGAGCTGA
Human LBP CDS (SEQ ID NO: 33) ATGGGGGCCTTGGCCAGAGCCCTGCCGTCCATACTGCTGGCATTGCTGCTTACGTCCACCCCAGAGG
CTCTGGGTGCCAACCCCGGCTTGGTCGCCAGGATCACCGACAAGGGACTGCAGTATGCGGCCCAGG
AGGGGCTATTAGCTCTGCAGAGTGAGCTGCTCAGGATCACGCTGCCTGACTTCACCGGGGACTTGA
GGATCCCCCACGTCGGCCGTGGGCGCTATGAGTTCCACAGCCTGAACATCCACAGCTGTGAGCTGC
TTCACTCTGCGCTGAGGCCTGTCCCTGGCCAGGGCCTGAGTCTCAGCATCTCCGACTCCTCCATCCG
GGTCCAGGGCAGGTGGAAGGTGCGCAAGTCATTCTTCAAACTACAGGGCTCCTTTGATGTCAGTGT
CAAGGGCATCAGCATTTCGGTCAACCTCCTGTTGGGCAGCGAGTCCTCCGGGAGGCCCACAGTTAC
TGCCTCCAGCTGCAGCAGTGACATCGCTGACGTGGAGGTGGACATGTCGGGAGACTTGGGGTGGCT
GTTGAACCTCTTCCACAACCAGATTGAGTCCAAGTTCCAGAAAGTACTGGAGAGCAGGATTTGCGA
AATGATCCAGAAATCAGTGTCCTCCGATCTACAGCCTTATCTCCAAACTCTGCCAGTTACAACAGAG
ATTGACAGTTTCGCCGACATTGATTATAGCTTAGTGGAAGCCCCTCGGGCAACAGCCCAGATGCTGG
AGGTGATGTTTAAGGGTGAAATCTTTCATCGTAACCACCGTTCTCCAGTTACCCTCCTTGCTGCAGTC
ATGAGCCTTCCTGAGGAACACAACAAAATGGTCTACTTTGCCATCTCGGATTATGTCTTCAACACGG
CCAGCCTGGTTTATCATGAGGAAGGATATCTGAACTTCTCCATCACAGATGACATGATACCGCCTGAC
TCTAATATCCGACTGACCACCAAGTCCTTCCGACCCTTCGTCCCACGGTTAGCCAGGCTCTACCCCA
ACATGAACCTGGAACTCCAGGGATCAGTGCCCTCTGCTCCGCTCCTGAACTTCAGCCCTGGGAATCT
GTCTGTGGACCCCTATATGGAGATAGATGCCTTTGTGCTCCTGCCCAGCTCCAGCAAGGAGCCTGTC
TTCCGGCTCAGTGTGGCCACTAATGTGTCCGCCACCTTGACCTTCAATACCAGCAAGATCACTGGGT
TCCTGAAGCCAGGAAAGGTAAAAGTGGAACTGAAAGAATCCAAAGTTGGACTATTCAATGCAGAG
CTGTTGGAAGCGCTCCTCAACTATTACATCCTTAACACCTTCTACCCCAAGTTCAATGATAAGTTGGC
CGAAGGCTTCCCCCTTCCTCTGCTGAAGCGTGTTCAGCTCTACGACCTTGGGCTGCAGATCCATAAG
GACTTCCTGTTCTTGGGTGCCAATGTCCAATACATGAGAGTTTGA
Human TRAF6 CDS (SEQ ID NO: 34) ATGAGTCTGCTAAACTGTGAAAACAGCTGTGGATCCAGCCAGTCTGAAAGTGACTGCTGTGTGGCC
ATGGCCAGCTCCTGTAGCGCTGTAACAAAAGATGATAGTGTGGGTGGAACTGCCAGCACGGGGAAC
CTCTCCAGCTCATTTATGGAGGAGATCCAGGGATATGATGTAGAGTTTGACCCACCCCTGGAAAGCA
AGTATGAATGCCCCATCTGCTTGATGGCATTACGAGAAGCAGTGCAAACGCCATGCGGCCATAGGTT
CTGCAAAGCCTGCATCATAAAATCAATAAGGGATGCAGGTCACAAATGTCCAGTTGACAATGAAATA
CTGCTGGAAAATCAACTATTTCCAGACAATTTTGCAAAACGTGAGATTCTTTCTCTGATGGTGAAAT
GTCCAAATGAAGGTTGTTTGCACAAGATGGAACTGAGACATCTTGAGGATCATCAAGCACATTGTG
AGTTTGCTCTTATGGATTGTCCCCAATGCCAGCGTCCCTTCCAAAAATTCCATATTAATATTCACATTC
TGAAGGATTGTCCAAGGAGACAGGTTTCTTGTGACAACTGTGCTGCATCAATGGCATTTGAAGATAA
AGAGATCCATGACCAGAACTGTCCTTTGGCAAATGTCATCTGTGAATACTGCAATACTATACTCATCA
GAGAACAGATGCCTAATCATTATGATCTAGACTGCCCTACAGCCCCAATTCCATGCACATTCAGTACT
TTTGGTTGCCATGAAAAGATGCAGAGGAATCACTTGGCACGCCACCTACAAGAGAACACCCAGTCA
CACATGAGAATGTTGGCCCAGGCTGTTCATAGTTTGAGCGTTATACCCGACTCTGGGTATATCTCAGA
GGTCCGGAATTTCCAGGAAACTATTCACCAGTTAGAGGGTCGCCTTGTAAGACAAGACCATCAAAT
CCGGGAGCTGACTGCTAAAATGGAAACTCAGAGTATGTATGTAAGTGAGCTCAAACGAACCATTCG
AACCCTTGAGGACAAAGTTGCTGAAATCGAAGCACAGCAGTGCAATGGAATTTATATTTGGAAGATT
GGCAACTTTGGAATGCATTTGAAATGTCAAGAAGAGGAGAAACCTGTTGTGATTCATAGCCCTGGAT
TCTACACTGGCAAACCCGGGTACAAACTGTGCATGCGCTTGCACCTTCAGTTACCGACTGCTCAGC
GCTGTGCAAACTATATATCCCTTTTTGTCCACACAATGCAAGGAGAATATGACAGCCACCTCCCTTGG
CCCTTCCAGGGTACAATACGCCTTACAATTCTTGATCAGTCTGAAGCACCTGTAAGGCAAAACCACG
AAGAGATAATGGATGCCAAACCAGAGCTGCTTGCTTTCCAGCGACCCACAATCCCACGGAACCCAA
AAGGTTTTGGCTATGTAACTTTTATGCATCTGGAAGCCCTAAGACAAAGAACTTTCATTAAGGATGA
CACATTATTAGTGCGCTGTGAGGTCTCCACCCGCTTTGACATGGGTAGCCTTCGGAGGGAGGGTTTT
CAGCCACGAAGTACTGATGCAGGGGTATAG

Human K-Ras CDS (SEQ ID NO: 35) ATGACTGAATATAAACTTGTGGTAGTTGGAGCTGGTGGCGTAGGCAAGAGTGCCTTGACGATACAGC
TAATTCAGAATCATTTTGTGGACGAATATGATCCAACAATAGAGGATTCCTACAGGAAGCAAGTAGT
AATTGATGGAGAAACCTGTCTCTTGGATATTCTCGACACAGCAGGTCAAGAGGAGTACAGTGCAAT
GAGGGACCAGTACATGAGGACTGGGGAGGGCTTTCTTTGTGTATTTGCCATAAATAATACTAAATCAT
TTGAAGATATTCACCATTATAGAGAACAAATTAAAAGAGTTAAGGACTCTGAAGATGTACCTATGGT
CCTAGTAGGAAATAAATGTGATTTGCCTTCTAGAACAGTAGACACAAAACAGGCTCAGGACTTAGC
AAGAAGTTATGGAATTCCTTTTATTGAAACATCAGCAAAGACAAGACAGGGTGTTGATGATGCCTTC
TATACATTAGTTCGAGAAATTCGAAAACATAAAGAAAAGATGAGCAAAGATGGTAAAAAGAAGAAA
AAGAAGTCAAAGACAAAGTGTGTAATTATGTAA
Human N-Ras CDS (SEQ ID NO: 36) ATGACTGAGTACAAACTGGTGGTGGTTGGAGCAGGTGGTGTTGGGAAAAGCGCACTGACAATCCA
GCTAATCCAGAACCACTTTGTAGATGAATATGATCCCACCATAGAGGATTCTTACAGAAAACAAGTG
GTTATAGATGGTGAAACCTGTTTGTTGGACATACTGGATACAGCTGGACAAGAAGAGTACAGTGCCA
TGAGAGACCAATACATGAGGACAGGCGAAGGCTTCCTCTGTGTATTTGCCATCAATAATAGCAAGTC
ATTTGCGGATATTAACCTCTACAGGGAGCAGATTAAGCGAGTAAAAGACTCGGATGATGTACCTATG
GTGCTAGTGGGAAACAAGTGTGATTTGCCAACAAGGACAGTTGATACAAAACAAGCCCACGAACT
GGCCAAGAGTTACGGGATTCCATTCATTGAAACCTCAGCCAAGACCAGACAGGGTGTTGAAGATGC
TTTTTACACACTGGTAAGAGAAATACGCCAGTACCGAATGAAAAAACTCAACAGCAGTGATGATGG
GACTCAGGGTTGTATGGGATTGCCATGTGTGGTGATGTAA
Human Raf CDS (SEQ ID NO: 37) ATGGCTAGCAAACGAAAATCTACAACTCCATGCATGGTTCGGACATCACAAGTAGTAGAACAAGAT
GTGCCCGAGGAAGTAGACAGGGCCAAAGAGAAAGGAATCGGCACACCACAGCCTGACGTGGCCA
AGGACAGTTGGGCAGCAGAACTTGAAAACTCTTCCAAAGAAAACGAAGTGATAGAGGTGAAATCT
ATGGGGGAAAGCCAGTCCAAAAAACTCCAAGGTGGTTATGAGTGCAAATACTGCCCCTACTCCACG
CAAAACCTGAACGAGTTCACGGAGCATGTCGACATGCAGCATCCCAACGTGATTCTCAACCCCCTC
TACGTGTGTGCAGAATGTAACTTCACAACCAAAAAGTACGACTCCCTATCCGACCACAACTCCAAG
TTCCATCCCGGGGAGGCCAACTTCAAGCTGAAGTTAATTAAACGCAATAATCAAACTGTCTTGGAAC
AGTCCATCGAAACCACCAACCATGTCGTGTCCATCACCACCAGTGGCCCTGGAACTGGTGACAGTG
ATTCTGGGATCTCGGTGAGTAAAACCCCCATCATGAAGCCTGGAAAACCAAAAGCGGATGCCAAGA
AGGTGCCCAAGAAGCCCGAGGAGATCACCCCCGAGAACCACGTGGAAGGGACCGCCCGCCTGGTG
ACAGACACAGCTGAGATCCTCTCGAGACTCGGCGGGGTGGAGCTCCTCCAAGACACATTAGGACA
CGTCATGCCTTCTGTACAGCTGCCACCAAATATCAACCTTGTGCCCAAGGTCCCTGTCCCACTAAATA
CTACCAAATACAACTCTGCCCTGGATACAAATGCCACGATGATCAACTCTTTCAACAAGTTTCCTTAC
CCGACCCAGGCTGAGTTGTCCTGGCTGACAGCTGCCTCCAAACACCCAGAGGAGCACATCAGAATC
TGGTTTGCCACCCAGCGCTTAAAGCATGGCATCAGCTGGTCCCCAGAAGAGGTGGAGGAGGCCCG
GAAGAAGATGTTCAACGGCACCATCCAGTCAGTACCCCCGACCATCACTGTGCTGCCCGCCCAGTT
GGCCCCCACAAAGGTGACGCAGCCCATCCTCCAGACGGCTCTACCGTGCCAGATCCTCGGCCAGAC
TAGCCTGGTGCTGACTCAGGTGACCAGCGGGTCAACAACCGTCTCTTGCTCCCCCATCACACTTGC
CGTGGCAGGAGTCACCAACCATGGCCAGAAGAGACCCTTGGTGACTCCCCAAGCTGCCCCCGAAC
CCAAGCGTCCACACATCGCTCAGGTGCCAGAGCCCCCACCCAAGGTGGCCAACCCCCCGCTCACAC
CAGCCAGTGACCGCAAGAAGACAAAGGAGCAGATAGCACATCTCAAGGCCAGCTTTCTCCAGAGC
CAGTTCCCTGACGATGCCGAGGTTTACCGGCTCATCGAGGTGACTGGCCTTGCCAGGAGCGAGATC
AAGAAGTGGTTCAGTGACCACCGATATCGGTGTCAAAGGGGCATCGTCCACATCACCAGCGAATCC
CTTGCCAAAGACCAGTTGGCCATCGCGGCCTCCCGACACGGTCGCACGTATCATGCGTACCCAGAC
TTTGCCCCCCAGAAGTTCAAAGAGAAAACACAGGGTCAGGTTAAAATCTTGGAAGACAGCTTTTTG
AAAAGTTCTTTTCCTACCCAAGCAGAACTGGATCGGCTAAGGGTGGAGACCAAGCTGAGCAGGAG
AGAGATCGACTCCTGGTTCTCGGAGAGGCGGAAGCTTCGAGACAGCATGGAACAAGCTGTCTTGG
ATTCCATGGGGTCTGGCAAAAAAGGCCAAGATGTGGGAGCCCCCAATGGTGCTCTGTCTCGACTCG
ACCAGCTCTCCGGTGCCCAGTTAACAAGTTCTCTGCCCAGCCCTTCGCCAGCAATTGCAAAAAGTC
AAGAACAGGTTCATCTCCTGAGGAGCACGTTTGCAAGAACCCAGTGGCCTACTCCCCAGGAGTACG

ACCAGTTAGCGGCCAAGACTGGCCTGGTCCGAACTGAGATTGTGCGTTGGTTCAAGGAGAACAGAT
GCTTGCTGAAAACGGGAACCGTGAAGTGGATGGAGCAGTACCAGCACCAGCCCATGGCAGATGAT
CACGGCTACGATGCCGTAGCAAGGAAAGCAACAAAACCCATGGCCGAGAGCCCAAAGAACGGGGG
TGATGTGGTTCCACAATATTACAAGGACCCCAAAAAGCTCTGCGAAGAGGACTTGGAGAAGTTGGT
GACCAGGGTAAAAGTAGGCAGCGAGCCAGCAAAAGACTGTTTGCCAGCAAAGCCCTCAGAGGCCA
CCTCAGACCGGTCAGAGGGCAGCAGCCGGGACGGCCAGGGTAGCGACGAGAACGAGGAGTCGAG
CGTTGTGGATTACGTGGAGGTGACGGTCGGGGAGGAGGATGCGATCTCAGATAGATCAGATAGCTG
GAGTCAGGCTGCGGCAGAAGGTGTGTCGGAACTGGCTGAATCAGACTCCGACTGCGTCCCTGCAG
AGGCTGGCCAGGCCTAG
Human MEK1 CDS (SEQ ID NO: 38) ATGCCCAAGAAGAAGCCGACGCCCATCCAGCTGAACCCGGCCCCCGACGGCTCTGCAGTTAACGG
GACCAGCTCTGCGGAGACCAACTTGGAGGCCTTGCAGAAGAAGCTGGAGGAGCTAGAGCTTGATG
AGCAGCAGCGAAAGCGCCTTGAGGCCTTTCTTACCCAGAAGCAGAAGGTGGGAGAACTGAAGGAT
GACGACTTTGAGAAGATCAGTGAGCTGGGGGCTGGCAATGGCGGTGTGGTGTTCAAGGTCTCCCAC
AAGCCTTCTGGCCTGGTCATGGCCAGAAAGCTAATTCATCTGGAGATCAAACCCGCAATCCGGAAC
CAGATCATAAGGGAGCTGCAGGTTCTGCATGAGTGCAACTCTCCGTACATCGTGGGCTTCTATGGTG
CGTTCTACAGCGATGGCGAGATCAGTATCTGCATGGAGCACATGGATGGAGGTTCTCTGGATCAAGT
CCTGAAGAAAGCTGGAAGAATTCCTGAACAAATTTTAGGAAAAGTTAGCATTGCTGTAATAAAAGG
CCTGACATATCTGAGGGAGAAGCACAAGATCATGCACAGAGATGTCAAGCCCTCCAACATCCTAGT
CAACTCCCGTGGGGAGATCAAGCTCTGTGACTTTGGGGTCAGCGGGCAGCTCATCGACTCCATGGC
CAACTCCTTCGTGGGCACAAGGTCCTACATGTCGCCAGAAAGACTCCAGGGGACTCATTACTCTGT
GCAGTCAGACATCTGGAGCATGGGACTGTCTCTGGTAGAGATGGCGGTTGGGAGGTATCCCATCCCT
CCTCCAGATGCCAAGGAGCTGGAGCTGATGTTTGGGTGCCAGGTGGAAGGAGATGCGGCTGAGAC
CCCACCCAGGCCAAGGACCCCCGGGAGGCCCCTTAGCTCATACGGAATGGACAGCCGACCTCCCAT
GGCAATTTTTGAGTTGTTGGATTACATAGTCAACGAGCCTCCTCCAAAACTGCCCAGTGGAGTGTTC
AGTCTGGAATTTCAAGATTTTGTGAATAAATGCTTAATAAAAAACCCCGCAGAGAGAGCAGATTTGA
AGCAACTCATGGTTCATGCTTTTATCAAGAGATCTGATGCTGAGGAAGTGGATTTTGCAGGTTGGCT
CTGCTCCACCATCGGCCTTAACCAGCCCAGCACACCAACCCATGCTGCTGGCGTCTAA
Human MEK2 CDS (SEQ ID NO: 39) ATGCTGGCCCGGAGGAAGCCGGTGCTGCCGGCGCTCACCATCAACCCTACCATCGCCGAGGGCCCA
TCCCCTACCAGCGAGGGCGCCTCCGAGGCAAACCTGGTGGACCTGCAGAAGAAGCTGGAGGAGCT
GGAACTTGACGAGCAGCAGAAGAAGCGGCTGGAAGCCTTTCTCACCCAGAAAGCCAAGGTCGGCG
AACTCAAAGACGATGACTTCGAAAGGATCTCAGAGCTGGGCGCGGGCAACGGCGGGGTGGTCACC
AAAGTCCAGCACAGACCCTCGGGCCTCATCATGGCCAGGAAGCTGATCCACCTTGAGATCAAGCCG
GCCATCCGGAACCAGATCATCCGCGAGCTGCAGGTCCTGCACGAATGCAACTCGCCGTACATCGTG
GGCTTCTACGGGGCCTTCTACAGTGACGGGGAGATCAGCATTTGCATGGAACACATGGACGGCGGC
TCCCTGGACCAGGTGCTGAAAGAGGCCAAGAGGATTCCCGAGGAGATCCTGGGGAAAGTCAGCAT
CGCGGTTCTCCGGGGCTTGGCGTACCTCCGAGAGAAGCACCAGATCATGCACCGAGATGTGAAGCC
CTCCAACATCCTCGTGAACTCTAGAGGGGAGATCAAGCTGTGTGACTTCGGGGTGAGCGGCCAGCT
CATCGACTCCATGGCCAACTCCTTCGTGGGCACGCGCTCCTACATGGCTCCGGAGCGGTTGCAGGG
CACACATTACTCGGTGCAGTCGGACATCTGGAGCATGGGCCTGTCCCTGGTGGAGCTGGCCGTCGG
AAGGTACCCCATCCCCCCGCCCGACGCCAAAGAGCTGGAGGCCATCTTTGGCCGGCCCGTGGTCGA
CGGGGAAGAAGGAGAGCCTCACAGCATCTCGCCTCGGCCGAGGCCCCCCGGGCGCCCCGTCAGCG
GTCACGGGATGGATAGCCGGCCTGCCATGGCCATCTTTGAACTCCTGGACTATATTGTGAACGAGCC
ACCTCCTAAGCTGCCCAACGGTGTGTTCACCCCCGACTTCCAGGAGTTTGTCAATAAATGCCTCATC
AAGAACCCAGCGGAGCGGGCGGACCTGAAGATGCTCACAAACCACACCTTCATCAAGCGGTCCGA
GGTGGAAGAAGTGGATTTTGCCGGCTGGTTGTGTAAAACCCTGCGGCTGAACCAGCCCGGCACACC
CACGCGCACCGCCGTGTGA
Human ERK1 CDS (SEQ ID NO: 40) ATGGCGGCGGCGGCGGCTCAGGGGGGCGGGGGCGGGGAGCCCCGTAGAACCGAGGGGGTCGGCC
CGGGGGTCCCGGGGGAGGTGGAGATGGTGAAGGGGCAGCCGTTCGACGTGGGCCCGCGCTACACG
CAGTTGCAGTACATCGGCGAGGGCGCGTACGGCATGGTCAGCTCGGCCTATGACCACGTGCGCAAG
ACTCGCGTGGCCATCAAGAAGATCAGCCCCTTCGAACATCAGACCTACTGCCAGCGCACGCTCCGG

GAGATCCAGATCCTGCTGCGCTTCCGCCATGAGAATGTCATCGGCATCCGAGACATTCTGCGGGCGT
CCACCCTGGAAGCCATGAGAGATGTCTACATTGTGCAGGACCTGATGGAGACTGACCTGTACAAGT
TGCTGAAAAGCCAGCAGCTGAGCAATGACCATATCTGCTACTTCCTCTACCAGATCCTGCGGGGCCT
CAAGTACATCCACTCCGCCAACGTGCTCCACCGAGATCTAAAGCCCTCCAACCTGCTCATCAACACC
ACCTGCGACCTTAAGATTTGTGATTTCGGCCTGGCCCGGATTGCCGATCCTGAGCATGACCACACCG
GCTTCCTGACGGAGTATGTGGCTACGCGCTGGTACCGGGCCCCAGAGATCATGCTGAACTCCAAGG
GCTATACCAAGTCCATCGACATCTGGTCTGTGGGCTGCATTCTGGCTGAGATGCTCTCTAACCGGCC
CATCTTCCCTGGCAAGCACTACCTGGATCAGCTCAACCACATTCTGGGCATCCTGGGCTCCCCATCC
CAGGAGGACCTGAATTGTATCATCAACATGAAGGCCCGAAACTACCTACAGTCTCTGCCCTCCAAG
ACCAAGGTGGCTTGGGCCAAGCTTTTCCCCAAGTCAGACTCCAAAGCCCTTGACCTGCTGGACCGG
ATGTTAACCTTTAACCCCAATAAACGGATCACAGTGGAGGAAGCGCTGGCTCACCCCTACCTGGAG
CAGTACTATGACCCGACGGATGAGGTGGGCCAGTCCCCAGCAGCAGTGGGGCTGGGGGCAGGGGA
GCAGGGGGGCACGTAG
Human ERK2 CDS (SEQ ID NO: 41) ATGGCGGCGGCGGCGGCGGCGGGCGCGGGCCCGGAGATGGTCCGCGGGCAGGTGTTCGACGTGGG
GCCGCGCTACACCAACCTCTCGTACATCGGCGAGGGCGCCTACGGCATGGTGTGCTCTGCTTATGAT
AATGTCAACAAAGTTCGAGTAGCTATCAAGAAAATCAGCCCCTTTGAGCACCAGACCTACTGCCAG
AGAACCCTGAGGGAGATAAAAATCTTACTGCGCTTCAGACATGAGAACATCATTGGAATCAATGAC
ATTATTCGAGCACCAACCATCGAGCAAATGAAAGATGTATATATAGTACAGGACCTCATGGAAACAG
ATCTTTACAAGCTCTTGAAGACACAACACCTCAGCAATGACCATATCTGCTATTTTCTCTACCAGATC
CTCAGAGGGTTAAAATATATCCATTCAGCTAACGTTCTGCACCGTGACCTCAAGCCTTCCAACCTGC
TGCTCAACACCACCTGTGATCTCAAGATCTGTGACTTTGGCCTGGCCCGTGTTGCAGATCCAGACCA
TGATCACACAGGGTTCCTGACAGAATATGTGGCCACACGTTGGTACAGGGCTCCAGAAATTATGTTG
AATTCCAAGGGCTACACCAAGTCCATTGATATTTGGTCTGTAGGCTGCATTCTGGCAGAAATGCTTTC
TAACAGGCCCATCTTTCCAGGGAAGCATTATCTTGACCAGCTGAACCACATTTTGGGTATTCTTGGAT
CCCCATCACAAGAAGACCTGAATTGTATAATAAATTTAAAAGCTAGGAACTATTTGCTTTCTCTTCCA
CACAAAAATAAGGTGCCATGGAACAGGCTGTTCCCAAATGCTGACTCCAAAGCTCTGGACTTATTG
GACAAAATGTTGACATTCAACCCACACAAGAGGATTGAAGTAGAACAGGCTCTGGCCCACCCATAT
CTGGAGCAGTATTACGACCCGAGTGACGAGCCCATCGCCGAAGCACCATTCAAGTTCGACATGGAA
TTGGATGACTTGCCTAAGGAAAAGCTCAAAGAACTAATTTTTGAAGAGACTGCTAGATTCCAGCCA
GGATACAGATCTTAA
Human IK13 CDS (SEQ ID NO: 42) ATGTTCCAGGCGGCCGAGCGCCCCCAGGAGTGGGCCATGGAGGGCCCCCGCGACGGGCTGAAGAA
GGAGCGGCTACTGGACGACCGCCACGACAGCGGCCTGGACTCCATGAAAGACGAGGAGTACGAGC
AGATGGTCAAGGAGCTGCAGGAGATCCGCCTCGAGCCGCAGGAGGTGCCGCGCGGCTCGGAGCCC
TGGAAGCAGCAGCTCACCGAGGACGGGGACTCGTTCCTGCACTTGGCCATCATCCATGAAGAAAAG
GCACTGACCATGGAAGTGATCCGCCAGGTGAAGGGAGACCTGGCCTTCCTCAACTTCCAGAACAAC
CTGCAGCAGACTCCACTCCACTTGGCTGTGATCACCAACCAGCCAGAAATTGCTGAGGCACTTCTG
GGAGCTGGCTGTGATCCTGAGCTCCGAGACTTTCGAGGAAATACCCCCCTACACCTTGCCTGTGAG
CAGGGCTGCCTGGCCAGCGTGGGAGTCCTGACTCAGTCCTGCACCACCCCGCACCTCCACTCCATC
CTGAAGGCTACCAACTACAATGGCCACACGTGTCTACACTTAGCCTCTATCCATGGCTACCTGGGCA
TCGTGGAGCTTTTGGTGTCCTTGGGTGCTGATGTCAATGCTCAGGAGCCCTGTAATGGCCGGACTGC
CCTTCACCTCGCAGTGGACCTGCAAAATCCTGACCTGGTGTCACTCCTGTTGAAGTGTGGGGCTGAT
GTCAACAGAGTTACCTACCAGGGCTATTCTCCCTACCAGCTCACCTGGGGCCGCCCAAGCACCCGG
ATACAGCAGCAGCTGGGCCAGCTGACACTAGAAAACCTTCAGATGCTGCCAGAGAGTGAGGATGA
GGAGAGCTATGACACAGAGTCAGAGTTCACGGAGTTCACAGAGGACGAGCTGCCCTATGATGACTG
TGTGTTTGGAGGCCAGCGTCTGACGTTATGA
Human Rac CDS (SEQ ID NO: 43) ATGAGCGACGTGGCTATTGTGAAGGAGGGTTGGCTGCACAAACGAGGGGAGTACATCAAGACCTG
GCGGCCACGCTACTTCCTCCTCAAGAATGATGGCACCTTCATTGGCTACAAGGAGCGGCCGCAGGA
TGTGGACCAACGTGAGGCTCCCCTCAACAACTTCTCTGTGGCGCAGTGCCAGCTGATGAAGACGGA

GCGGCCCCGGCCCAACACCTTCATCATCCGCTGCCTGCAGTGGACCACTGTCATCGAACGCACCTTC
CATGTGGAGACTCCTGAGGAGCGGGAGGAGTGGACAACCGCCATCCAGACTGTGGCTGACGGCCT
CAAGAAGCAGGAGGAGGAGGAGATGGACTTCCGGTCGGGCTCACCCAGTGACAACTCAGGGGCTG
AAGAGATGGAGGTGTCCCTGGCCAAGCCCAAGCACCGCGTGACCATGAACGAGTTTGAGTACCTG
AAGCTGCTGGGCAAGGGCACTTTCGGCAAGGTGATCCTGGTGAAGGAGAAGGCCACAGGCCGCTA
CTACGCCATGAAGATCCTCAAGAAGGAAGTCATCGTGGCCAAGGACGAGGTGGCCCACACACTCA
CCGAGAACCGCGTCCTGCAGAACTCCAGGCACCCCTTCCTCACAGCCCTGAAGTACTCTTTCCAGA
CCCACGACCGCCTCTGCTTTGTCATGGAGTACGCCAACGGGGGCGAGCTGTTCTTCCACCTGTCCC
GGGAGCGTGTGTTCTCCGAGGACCGGGCCCGCTTCTATGGCGCTGAGATTGTGTCAGCCCTGGACT
ACCTGCACTCGGAGAAGAACGTGGTGTACCGGGACCTCAAGCTGGAGAACCTCATGCTGGACAAG
GACGGGCACATTAAGATCACAGACTTCGGGCTGTGCAAGGAGGGGATCAAGGACGGTGCCACCAT
GAAGACCTTTTGCGGCACACCTGAGTACCTGGCCCCCGAGGTGCTGGAGGACAATGACTACGGCCG
TGCAGTGGACTGGTGGGGGCTGGGCGTGGTCATGTACGAGATGATGTGCGGTCGCCTGCCCTTCTA
CAACCAGGACCATGAGAAGCTTTTTGAGCTCATCCTCATGGAGGAGATCCGCTTCCCGCGCACGCTT
GGTCCCGAGGCCAAGTCCTTGCTTTCAGGGCTGCTCAAGAAGGACCCCAAGCAGAGGCTTGGCGG
GGGCTCCGAGGACGCCAAGGAGATCATGCAGCATCGCTTCTTTGCCGGTATCGTGTGGCAGCACGT
GTACGAGAAGAAGCTCAGCCCACCCTTCAAGCCCCAGGTCACGTCGGAGACTGACACCAGGTATTT
TGATGAGGAGTTCACGGCCCAGATGATCACCATCACACCACCTGACCAAGATGACAGCATGGAGTG
TGTGGACAGCGAGCGCAGGCCCCACTTCCCCCAGTTCTCCTACTCGGCCAGCGGCACGGCCTGA
Human MEK3 CDS (SEQ ID NO: 44) ATGTCCAAGCCACCCGCACCCAACCCCACACCCCCCCGGAACCTGGACTCCCGGACCTTCATCACC
ATTGGAGACAGAAACTTTGAGGTGGAGGCTGATGACTTGGTGACCATCTCAGAACTGGGCCGTGGA
GCCTATGGGGTGGTAGAGAAGGTGCGGCACGCCCAGAGCGGCACCATCATGGCCGTGAAGCGGATC
CGGGCCACCGTGAACTCACAGGAGCAGAAGCGGCTGCTCATGGACCTGGACATCAACATGCGCAC
GGTCGACTGTTTCTACACTGTCACCTTCTACGGGGCACTATTCAGAGAGGGAGACGTGTGGATCTGC
ATGGAGCTCATGGACACATCCTTGGACAAGTTCTACCGGAAGGTGCTGGATAAAAACATGACAATTC
CAGAGGACATCCTTGGGGAGATTGCTGTGTCTATCGTGCGGGCCCTGGAGCATCTGCACAGCAAGC
TGTCGGTGATCCACAGAGATGTGAAGCCCTCCAATGTCCTTATCAACAAGGAGGGCCATGTGAAGA
TGTGTGACTTTGGCATCAGTGGCTACTTGGTGGACTCTGTGGCCAAGACGATGGATGCCGGCTGCA
AGCCCTACATGGCCCCTGAGAGGATCAACCCAGAGCTGAACCAGAAGGGCTACAATGTCAAGTCCG
ACGTCTGGAGCCTGGGCATCACCATGATTGAGATGGCCATCCTGCGGTTCCCTTACGAGTCCTGGGG
GACCCCGTTCCAGCAGCTGAAGCAGGTGGTGGAGGAGCCGTCCCCCCAGCTCCCAGCCGACCGTT
TCTCCCCCGAGTTTGTGGACTTCACTGCTCAGTGCCTGAGGAAGAACCCCGCAGAGCGTATGAGCT
ACCTGGAGCTGATGGAGCACCCCTTCTTCACCTTGCACAAAACCAAGAAGACGGACATTGCTGCCT
TCGTGAAGGAGATCCTGGGAGAAGACTCATAG
Human MEK6 CDS (SEQ ID NO: 45) ATGGAACTGGGACGAGGTGCGTACGGGGTGGTGGAGAAGATGCGGCACGTGCCCAGCGGGCAGAT
CATGGCAGTGAAGCGGATCCGAGCCACAGTAAATAGCCAGGAACAGAAACGGCTACTGATGGATTT
GGATATTTCCATGAGGACGGTGGACTGTCCATTCACTGTCACCTTTTATGGCGCACTGTTTCGGGAG
GGTGATGTGTGGATCTGCATGGAGCTCATGGATACATCACTAGATAAATTCTACAAACAAGTTATTGA
TAAAGGCCAGACAATTCCAGAGGACATCTTAGGGAAAATAGCAGTTTCTATTGTAAAAGCATTAGAA
CATTTACATAGTAAGCTGTCTGTCATTCACAGAGACGTCAAGCCTTCTAATGTACTCATCAATGCTCT
CGGTCAAGTGAAGATGTGCGATTTTGGAATCAGTGGCTACTTGGTGGACTCTGTTGCTAAAACAATT
GATGCAGGTTGCAAACCATACATGGCCCCTGAAAGAATAAACCCAGAGCTCAACCAGAAGGGATAC
AGTGTGAAGTCTGACATTTGGAGTCTGGGCATCACGATGATTGAGTTGGCCATCCTTCGATTTCCCT
ATGATTCATGGGGAACTCCATTTCAGCAGCTCAAACAGGTGGTAGAGGAGCCATCGCCACAACTCC
CAGCAGACAAGTTCTCTGCAGAGTTTGTTGACTTTACCTCACAGTGCTTAAAGAAGAATTCCAAAG
AACGGCCTACATACCCAGAGCTAATGCAACATCCATTTTTCACCCTACATGAATCCAAAGGAACAGA
TGTGGCATCTTTTGTAAAACTGATTCTTGGAGACTAA
Human p38 CDS (SEQ ID NO: 46) ATGTCTCAGGAGAGGCCCACGTTCTACCGGCAGGAGCTGAACAAGACAATCTGGGAGGTGCCCGA
GCGTTACCAGAACCTGTCTCCAGTGGGCTCTGGCGCCTATGGCTCTGTGTGTGCTGCTTTTGACACA

AAAACGGGGTTACGTGTGGCAGTGAAGAAGCTCTCCAGACCATTTCAGTCCATCATTCATGCGAAA
AGAACCTACAGAGAACTGCGGTTACTTAAACATATGAAACATGAAAATGTGATTGGTCTGTTGGACG
TTTTTACACCTGCAAGGTCTCTGGAGGAATTCAATGATGTGTATCTGGTGACCCATCTCATGGGGGC
AGATCTGAACAACATTGTGAAATGTCAGAAGCTTACAGATGACCATGTTCAGTTCCTTATCTACCAA
ATTCTCCGAGGTCTAAAGTATATACATTCAGCTGACATAATTCACAGGGACCTAAAACCTAGTAATCT
AGCTGTGAATGAAGACTGTGAGCTGAAGATTCTGGATTTTGGACTGGCTCGGCACACAGATGATGA
AATGACAGGCTACGTGGCCACTAGGTGGTACAGGGCTCCTGAGATCATGCTGAACTGGATGCATTAC
AACCAGACAGTTGATATTTGGTCAGTGGGATGCATAATGGCCGAGCTGTTGACTGGAAGAACATTGT
TTCCTGGTACAGACCATATTAACCAGCTTCAGCAGATTATGCGTCTGACAGGAACACCCCCCGCTTAT
CTCATTAACAGGATGCCAAGCCATGAGGCAAGAAACTATATTCAGTCTTTGACTCAGATGCCGAAGA
TGAACTTTGCGAATGTATTTATTGGTGCCAATCCCCTGGCTGTCGACTTGCTGGAGAAGATGCTTGTA
TTGGACTCAGATAAGAGAATTACAGCGGCCCAAGCCCTTGCACATGCCTACTTTGCTCAGTACCACG
ATCCTGATGATGAACCAGTGGCCGATCCTTATGATCAGTCCTTTGAAAGCAGGGACCTCCTTATAGAT
GAGTGGAAAAGCCTGACCTATGATGAAGTCATCAGCTTTGTGCCACCACCCCTTGACCAAGAAGAG
ATGGAGTCCTGA
Human PKR CDS (SEQ ID NO: 47) ATGGCTGGTGATCTTTCAGCAGGTTTCTTCATGGAGGAACTTAATACATACCGTCAGAAGCAGGGAG
TAGTACTTAAATATCAAGAACTGCCTAATTCAGGACCTCCACATGATAGGAGGTTTACATTTCAAGTT
ATAATAGATGGAAGAGAATTTCCAGAAGGTGAAGGTAGATCAAAGAAGGAAGCAAAAAATGCCGC
AGCCAAATTAGCTGTTGAGATACTTAATAAGGAAAAGAAGGCAGTTAGTCCTTTATTATTGACAACA
ACGAATTCTTCAGAAGGATTATCCATGGGGAATTACATAGGCCTTATCAATAGAATTGCCCAGAAGA
AAAGACTAACTGTAAATTATGAACAGTGTGCATCGGGGGTGCATGGGCCAGAAGGATTTCATTATAA
ATGCAAAATGGGACAGAAAGAATATAGTATTGGTACAGGTTCTACTAAACAGGAAGCAAAACAATT
GGCCGCTAAACTTGCATATCTTCAGATATTATCAGAAGAAACCTCAGTGAAATCTGACTACCTGTCCT
CTGGTTCTTTTGCTACTACGTGTGAGTCCCAAAGCAACTCTTTAGTGACCAGCACACTCGCTTCTGA
ATCATCATCTGAAGGTGACTTCTCAGCAGATACATCAGAGATAAATTCTAACAGTGACAGTTTAAAC
AGTTCTTCGTTGCTTATGAATGGTCTCAGAAATAATCAAAGGAAGGCAAAAAGATCTTTGGCACCCA
GATTTGACCTTCCTGACATGAAAGAAACAAAGTATACTGTGGACAAGAGGTTTGGCATGGATTTTAA
AGAAATAGAATTAATTGGCTCAGGTGGATTTGGCCAAGTTTTCAAAGCAAAACACAGAATTGACGG
AAAGACTTACGTTATTAAACGTGTTAAATATAATAACGAGAAGGCGGAGCGTGAAGTAAAAGCATTG
GCAAAACTTGATCATGTAAATATTGTTCACTACAATGGCTGTTGGGATGGATTTGATTATGATCCTGA
GACCAGTGATGATTCTCTTGAGAGCAGTGATTATGATCCTGAGAACAGCAAAAATAGTTCAAGGTCA
AAGACTAAGTGCCTTTTCATCCAAATGGAATTCTGTGATAAAGGGACCTTGGAACAATGGATTGAAA
AAAGAAGAGGCGAGAAACTAGACAAAGTTTTGGCTTTGGAACTCTTTGAACAAATAACAAAAGGG
GTGGATTATATACATTCAAAAAAATTAATTCATAGAGATCTTAAGCCAAGTAATATATTCTTAGTAGAT
ACAAAACAAGTAAAGATTGGAGACTTTGGACTTGTAACATCTCTGAAAAATGATGGAAAGCGAACA
AGGAGTAAGGGAACTTTGCGATACATGAGCCCAGAACAGATTTCTTCGCAAGACTATGGAAAGGAA
GTGGACCTCTACGCTTTGGGGCTAATTCTTGCTGAACTTCTTCATGTATGTGACACTGCTTTTGAAAC
ATCAAAGTTTTTCACAGACCTACGGGATGGCATCATCTCAGATATATTTGATAAAAAAGAAAAAACT
CTTCTACAGAAATTACTCTCAAAGAAACCTGAGGATCGACCTAACACATCTGAAATACTAAGGACCT
TGACTGTGTGGAAGAAAAGCCCAGAGAAAAATGAACGACACACATGTTAG
Human TTP CDS (SEQ ID NO: 48) ATGGCGGCTCAGCGGATCCGAGCGGCCAACTCCAATGGCCTCCCTCGCTGCAAGTCAGAGGGGACC
CTGATTGACCTGAGCGAAGGGTTTTCAGAGACGAGCTTTAATGACATCAAAGTGCCTTCTCCCAGT
GCCTTGCTCGTAGACAACCCCACACCTTTCGGAAATGCAAAGGAAGTGATTGCGATCAAGGACTAT
TGCCCCACCAACTTCACCACACTGAAGTTCTCCAAGGGCGACCATCTCTACGTCTTGGACACATCTG
GCGGTGAGTGGTGGTACGCACACAACACCACCGAAATGGGCTACATCCCCTCCTCCTATGTGCAGC
CCTTGAACTACCGGAACTCAACACTGAGTGACAGCGGTATGATTGATAATCTTCCAGACAGCCCAG
ACGAGGTAGCCAAGGAGCTGGAGCTGCTCGGGGGATGGACAGATGACAAAAAAGTACCAGGCAGA
ATGTACAGTAATAACCCTTTCTGGAATGGGGTCCAGACCAATCCATTTCTGAATGGGAACGTGCCCG
TCATGCCCAGCCTGGATGAGCTGAATCCCAAAAGTACTGTGGATTTGCTCCTTTTTGACGCAGGTAC
ATCCTCCTTCACCGAATCCAGCTCAGCCACCACGAATAGCACTGGCAACATCTTCGATGAGCTTCCA
GTCACAAACGGACTCCACGCAGAGCCGCCGGTCAGGCGGGACAACCCCTTCTTCAGAAGCAAGCG
CTCCTACAGTCTCTCGGAACTCTCCGTCCTCCAAGCCAAGTCCGATGCTCCCACATCGTCGAGTTTC

TTCACCGGCTTGAAATCACCTGCCCCCGAGCAATTTCAGAGCCGGGAGGATTTTCGAACTGCCTGG
CTAAACCACAGGAAGCTGGCCCGGTCTTGCCACGACCTGGACTTGCTTGGCCAAAGCCCTGGTTGG
GGCCAGACCCAAGCCGTGGAGACAAACATCGTGTGCAAGCTGGATAGCTCCGGGGGTGCTGTCCA
GCTTCCTGACACCAGCATCAGCATCCACGTGCCCGAGGGCCACGTCGCCCCTGGGGAGACCCAGCA
GATCTCCATGAAAGCCCTGCTGGACCCCCCGCTGGAGCTCAACAGTGACAGGTCCTGCAGCATCAG
CCCTGTGCTGGAGGTCAAGCTGAGCAACCTGGAGGTGAAAACCTCTATCATCTTGGAGATGAAAGT
GTCAGCCGAGATAAAAAATGACCTTTTTAGCAAAAGCACAGTGGGCCTCCAGTGCCTGAGGAGCGA
CTCGAAGGAAGGGCCATATGTCTCCGTCCCGCTCAACTGCAGCTGTGGGGACACGGTCCAGGCACA
GCTGCACAACCTGGAGCCCTGTATGTACGTGGCTGTCGTGGCCCATGGCCCAAGCATCCTCTACCCT
TCCACCGTGTGGGACTTCATCAATAAAAAAGTCACAGTGGGTCTCTACGGCCCTAAACACATCCACC
CATCCTTCAAGACGGTAGTGACCATTTTTGGGCATGACTGTGCCCCAAAGACGCTCCTGGTCAGCG
AGGTCACACGCCAGGCACCCAACCCTGCCCCGGTGGCCCTGCAGCTGTGGGGGAAGCACCAGTTC
GTTTTGTCCAGGCCCCAGGATCTCAAGGTCTGTATGTTTTCCAATATGACGAATTACGAGGTCAAAG
CCAGCGAGCAGGCCAAAGTGGTGCGAGGATTCCAGCTGAAGCTGGGCAAGGTGAGCCGCCTGATC
TTCCCCATCACCTCCCAGAACCCCAACGAGCTCTCTGACTTCACGCTGCGGGTTCAGGTGAAGGAC
GACCAGGAGGCCATCCTCACCCAGTTTTGTGTCCAGACTCCTCAGCCACCCCCTAAAAGTGCCATC
AAGCCTTCCGGGCAAAGGAGGTTTCTCAAGAAGAACGAAGTCGGGAAAATCATCCTGTCCCCGTTT
GCCACCACTACAAAGTACCCGACTTTCCAGGACCGCCCGGTGTCCAGCCTCAAGTTTGGTAAGTTG
CTCAAGACTGTGGTGCGGCAGAACAAGAACCACTACCTGCTGGAGTACAAGAAGGGCGACGGGAT
CGCCCTGCTCAGCGAGGAGCGGGTCAGGCTCCGGGGCCAGCTGTGGACCAAGGAGTGGTACATCG
GCTACTACCAGGGCAGGGTGGGCCTCGTGCACACCAAGAACGTGCTGGTGGTCGGCAGGGCCCGG
CCCAGCCTGTGCTCGGGCCCCGAGCTGAGCACCTCGGTGCTGCTGGAGCAGATCCTGCGGCCCTGC
AAATTCCTCACGTACATCTATGCCTCCGTGAGGACCCTGCTCATGGAGAACATCAGCAGCTGGCGCT
CCTTCGCTGACGCCCTGGGCTACGTGAACCTGCCGCTCACCTTTTTCTGCCGGGCAGAGCTGGATAG
TGAGCCCGAGCGGGTGGCGTCCGTCCTAGAAAAGCTGAAGGAGGACTGTAACAACACTGAGAACA
AAGAACGGAAGTCCTTCCAGAAGGAGCTTGTGATGGCCCTACTGAAGATGGACTGCCAGGGCCTG
GTGGTCAGACTCATCCAGGACTTTGTGCTCCTGACCACGGCTGTAGAGGTGGCCCAGCGCTGGCGG
GAGCTGGCTGAGAAGCTGGCCAAGGTCTCCAAGCAGCAGATGGACGCCTACGAGTCTCCCCACCG
GGACAGGAACGGGGTTGTGGACAGCGAGGCCATGTGGAAGCCTGCGTATGACTTCTTACTCACCTG
GAGCCATCAGATCGGGGACAGCTACCGGGATGTCATCCAGGAGCTGCACCTGGGCCTGGACAAGAT
GAAAAACCCCATCACCAAGCGCTGGAAGCACCTCACTGGGACTCTGATCTTGGTGAACTCCCTGGA
CGTTCTGAGAGCAGCCGCCTTCAGCCCTGCGGACCAGGACGACTTCGTGATTTGA
Human MK2 CDS (SEQ ID NO: 49) ATGCTGTCCAACTCCCAGGGCCAGAGCCCGCCGGTGCCGTTCCCCGCCCCGGCCCCGCCGCCGCAG
CCCCCCACCCCTGCCCTGCCGCACCCCCCGGCGCAGCCGCCGCCGCCGCCCCCGCAGCAGTTCCCG
CAGTTCCACGTCAAGTCCGGCCTGCAGATCAAGAAGAACGCCATCATCGATGACTACAAGGTCACC
AGCCAGGTCCTGGGGCTGGGCATCAACGGCAAAGTTTTGCAGATCTTCAACAAGAGGACCCAGGA
GAAATTCGCCCTCAAAATGCTTCAGGACTGCCCCAAGGCCCGCAGGGAGGTGGAGCTGCACTGGC
GGGCCTCCCAGTGCCCGCACATCGTACGGATCGTGGATGTGTACGAGAATCTGTACGCAGGGAGGA
AGTGCCTGCTGATTGTCATGGAATGTTTGGACGGTGGAGAACTCTTTAGCCGAATCCAGGATCGAGG
AGACCAGGCATTCACAGAAAGAGAAGCATCCGAAATCATGAAGAGCATCGGTGAGGCCATCCAGTA
TCTGCATTCAATCAACATTGCCCATCGGGATGTCAAGCCTGAGAATCTCTTATACACCTCCAAAAGG
CCCAACGCCATCCTGAAACTCACTGACTTTGGCTTTGCCAAGGAAACCACCAGCCACAACTCTTTG
ACCACTCCTTGTTATACACCGTACTATGTGGCTCCAGAAGTGCTGGGTCCAGAGAAGTATGACAAGT
CCTGTGACATGTGGTCCCTGGGTGTCATCATGTACATCCTGCTGTGTGGGTATCCCCCCTTCTACTCC
AACCACGGCCTTGCCATCTCTCCGGGCATGAAGACTCGCATCCGAATGGGCCAGTATGAATTTCCCA
ACCCAGAATGGTCAGAAGTATCAGAGGAAGTGAAGATGCTCATTCGGAATCTGCTGAAAACAGAGC
CCACCCAGAGAATGACCATCACCGAGTTTATGAACCACCCTTGGATCATGCAATCAACAAAGGTCCC
TCAAACCCCACTGCACACCAGCCGGGTCCTGAAGGAGGACAAGGAGCGGTGGGAGGATGTCAAGG
GGTGTCTTCATGACAAGAACAGCGACCAGGCCACTTGGCTGACCAGGTTGTGA
An antisense nucleic acid molecule can be complementary to all or part of a non-coding region of the coding strand of a nucleotide sequence encoding a TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 protein. Non-coding regions (5' and 3' untranslated regions) are the 5' and 3' sequences that flank the coding region in a gene and are not translated into amino acids.
Based upon the sequences disclosed herein, one of skill in the art can easily choose and synthesize any of a number of appropriate antisense nucleic acids to target a nucleic acid encoding a TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 protein described herein. Antisense nucleic acids targeting a nucleic acid encoding a TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 protein can be designed using the software available at the Integrated DNA Technologies website.
An antisense nucleic acid can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides or more in length. An antisense oligonucleotide can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used.
Examples of modified nucleotides which can be used to generate an antisense nucleic acid include 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethy1-2-thiouracil, beta-D-mannosylqueosine, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methy1-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methy1-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest).
The antisense nucleic acid molecules described herein can be prepared in vitro and administered to a mammal, e.g., a human. Alternatively, they can be generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding a TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK
3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, LBP, TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 protein to thereby inhibit expression, e.g., by inhibiting transcription and/or translation.
The hybridization can be by conventional nucleotide complementarities to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA
duplexes, through specific interactions in the major groove of the double helix. The antisense nucleic acid molecules can be delivered to a mammalian cell using a vector (e.g., a lentivirus, a retrovirus, or an adenovirus vector).
An antisense nucleic acid can be an a-anomeric nucleic acid molecule. An a-anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual, 13-units, the strands run parallel to each other (Gaultier et al., Nucleic Acids Res. 15:6625-6641, 1987). The antisense nucleic acid can also comprise a 2'-0-methylribonucleotide (Inoue et al., Nucleic Acids Res. 15:6131-6148, 1987) or a chimeric RNA-DNA analog (Inoue et al., FEBS Lett. 215:327-330, 1987).
Another example of an inhibitory nucleic acid is a ribozyme that has specificity for a nucleic acid encoding a TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, INK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 protein (e.g., specificity for a TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-I, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, INK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA, e.g., specificity for any one of SEQ ID NOs: 13-49). Ribozymes are catalytic RNA
molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes (described in Haselhoff and Gerlach, Nature 334:585-591, 1988)) can be used to catalytically cleave mRNA transcripts to thereby inhibit translation of the protein encoded by the mRNA. A ribozyme having specificity for a TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, INK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA can be designed based upon the nucleotide sequence of any of the TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK
3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA sequences disclosed herein. For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in a TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA (see, e.g., U.S. Patent. Nos.
4,987,071 and 5,116,742). Alternatively, a TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, INK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA
can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA
molecules. See, e.g., Bartel et al., Science 261:1411-1418, 1993.
An inhibitory nucleic acid can also be a nucleic acid molecule that forms triple helical structures. For example, expression of a TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, INK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, INK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 polypeptide can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the gene encoding the TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, INK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 polypeptide (e.g., the promoter and/or enhancer, e.g., a sequence that is at least 1 kb, 2 kb, 3 kb, 4 kb, or 5 kb upstream of the transcription initiation start state) to form triple helical structures that prevent transcription of the gene in target cells. See generally Helene, Anticancer Drug Des. 6(6):569-84, 1991; Helene, Ann. N.Y. Acad. Sci. 660:27-36, 1992; and Maher, Bioassays 14(12):807-15, 1992.
In various embodiments, inhibitory nucleic acids can be modified at the base moiety, sugar moiety, or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acids can be modified to generate peptide nucleic acids (see, e.g., Hyrup et al., Bioorganic Medicinal Chem. 4(1):5-23, 1996). Peptide nucleic acids (PNAs) are nucleic acid mimics, e.g., DNA mimics, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs allows for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols (see, e.g., Perry-O'Keefe et al., Proc.
Natl. Acad. Sci.
U.S.A. 93:14670-675, 1996). PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication.
PNAs can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA
chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras can be generated which may combine the advantageous properties of PNA
and DNA. Such chimeras allow DNA recognition enzymes, e.g., RNAse H and DNA
polymerases, to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation.

The synthesis of PNA-DNA chimeras can be performed as described in Finn et al., Nucleic Acids Res. 24:3357-63, 1996. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry and modified nucleoside analogs. Compounds such as 5'-(4-methoxytrityl)amino-5'-deoxy-thymidine phosphoramidite can be used as a link between the PNA and the 5' end of DNA
(Mag et al., Nucleic Acids Res. 17:5973-88, 1989). PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5' PNA segment and a 3' DNA
segment (Finn et al., Nucleic Acids Res. 24:3357-63, 1996). Alternatively, chimeric molecules can be synthesized with a 5' DNA segment and a 3' PNA segment (Peterser et al., Bioorganic Med.
Chem. Lett. 5:1119-11124, 1975).
In some embodiments, the inhibitory nucleic acids can include other appended groups such as peptides, or agents facilitating transport across the cell membrane (see, Letsinger et al., Proc. Natl. Acad. Sci. U.S.A. 86:6553-6556, 1989; Lemaitre et al., Proc.
Natl. Acad. Sci.
U.S.A. 84:648-652, 1989; and WO 88/09810). In addition, the inhibitory nucleic acids can be modified with hybridization-triggered cleavage agents (see, e.g., Krol et al., Bio/Techniques 6:958-976, 1988) or intercalating agents (see, e.g., Zon, Pharm. Res., 5:539-549, 1988). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent, etc.
Another means by which expression of a TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, INK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA can be decreased in a mammalian cell is by RNA interference (RNAi).
RNAi is a process in which mRNA is degraded in host cells. To inhibit an mRNA, double-stranded RNA (dsRNA) corresponding to a portion of the gene to be silenced (e.g., a gene encoding a TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, fkB, NF-KB, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 polypeptide) is introduced into a mammalian cell. The dsRNA is digested into 21-23 nucleotide-long duplexes called short interfering RNAs (or siRNAs), which bind to a nuclease complex to form what is known as the RNA-induced silencing complex (or RISC). The RISC targets the homologous transcript by base pairing interactions between one of the siRNA strands and the endogenous mRNA.
It then cleaves the mRNA about 12 nucleotides from the 3' terminus of the siRNA (see Sharp et al., Genes Dev. 15:485-490, 2001, and Hammond et al., Nature Rev. Gen. 2:110-119, 2001).
RNA-mediated gene silencing can be induced in a mammalian cell in many ways, e.g., by enforcing endogenous expression of RNA hairpins (see, Paddison et al., Proc. Natl.
Acad. Sci. U.S.A. 99:1443-1448, 2002) or, as noted above, by transfection of small (21-23 nt) dsRNA (reviewed in Caplen, Trends Biotech. 20:49-51, 2002). Methods for modulating gene expression with RNAi are described, e.g., in U.S. Patent No. 6,506,559 and US
2003/0056235, which are hereby incorporated by reference.
Standard molecular biology techniques can be used to generate siRNAs. Short interfering RNAs can be chemically synthesized, recombinantly produced, e.g., by expressing RNA from a template DNA, such as a plasmid, or obtained from commercial vendors, such as Dharmacon. The RNA used to mediate RNAi can include synthetic or modified nucleotides, such as phosphorothioate nucleotides. Methods of transfecting cells with siRNA
or with plasmids engineered to make siRNA are routine in the art.
The siRNA molecules used to decrease expression of a TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, fkB, NF-KB, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA can vary in a number of ways. For example, they can include a 3' hydroxyl group and strands of 21, 22, or 23 consecutive nucleotides. They can be blunt ended or include an overhanging end at either the 3' end, the 5' end, or both ends. For example, at least one strand of the RNA molecule can have a 3' overhang from about 1 to about 6 nucleotides (e.g., 1-5, 1-3, 2-4, or 3-5 nucleotides (whether pyrimidine or purine nucleotides) in length. Where both strands include an overhang, the length of the overhangs may be the same or different for each strand.
To further enhance the stability of the RNA duplexes, the 3' overhangs can be stabilized against degradation (by, e.g., including purine nucleotides, such as adenosine or guanosine nucleotides or replacing pyrimidine nucleotides by modified analogues (e.g., substitution of uridine 2-nucleotide 3' overhangs by 2'-deoxythymidine is tolerated and does not affect the efficiency of RNAi). Any siRNA can be used in the methods of decreasing a TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, INK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 mRNA, provided it has sufficient homology to the target of interest (e.g., a sequence present in any one of SEQ ID NOs: 13-49, e.g., a target sequence encompassing the translation start site or the first exon of the mRNA). There is no upper limit on the length of the siRNA that can be used (e.g., the siRNA can range from about 21 base pairs of the gene to the full length of the gene or more (e.g., about 20 to about 30 base pairs, about 50 to about 60 base pairs, about 60 to about 70 base pairs, about 70 to about 80 base pairs, about 80 to about 90 base pairs, or about 90 to about 100 base pairs).
Exemplary TNFa inhibitors that are inhibitory nucleic acids targeting TNFa include, e.g., antisense DNA (e.g., Myers et al., J Pharmacol Exp Ther. 304(1):411-424, 2003;
Wasmuth et al., Invest. Opthalmol. Vis. Sci, 2003; Dong et al., J. Orthop.
Res. 26(8):1114-1120, 2008; U.S. Patent Application Serial Nos. 2003/0083275, 2003/0022848, and 2004/0770970; ISIS 104838; U.S. Patent Nos. 6,180,403, 6,080,580, and 6,228,642; Kobzik et al., Inhibition of TNF Synthesis by Antisense Oligonucleotides, in Manual of Antisense Methodology, Kluwer Academic Publishers, Vol. 4, pp.107-123, 1999; Taylor et al., Antisense Nucleic Acid Drug Develop. 8(3):199-205, 1998; Mayne et al., Stroke 32:240-248, 2001; Mochizuki et al., J. Controlled Release 151(2):155-161, 2011; Dong et al., J.
Orthopaedic Res. 26(8):1114-1120, 2008; Dong et al., Pharm. Res. 28(6):1349-1356, 2011;
and Pampfer et al., Biol. Reproduction 52(6):1316-1326, 1995), antisense RNA, short interfering RNA (siRNA) (e.g., Taishi et al., Brain Research 1156:125-132, 2007; Presumey et al., Eur. I Pharm. Biopharm. 82(3):457-467, 2012; Laroui et al., J.
Controlled Release 186:41-53, 2014; D'Amore et al., Int. J. Immunopathology Pharmacol. 21:1045-1047, 2008;
Choi et al., J. Dermatol. Sci. 52:87-97, 2008; Qin et al., Artificial Organs 35:706-714, 2011;
McCarthy et al., J. Controlled Release 168: 28-34, 2013; Khoury et al., Current Opin. Mol.
Therapeutics 9(5):483-489, 2007; Lu et al., RNA Interference Technology From Basic Science to Drug Development 303, 2005; Xie et al., PharmaGenomics 4(6):28-34, 2004;
Aldawsari et al., Current Pharmaceutical Design 21(31):4594-4605, 2015; Zheng et al., Arch.
Med. Sci.

11:1296-1302, 2015; Peng et al., Chinese J. Surgery 47(5):377-380, 2009;
Aldayel et al., Molecular Therapy. Nucleic Acids 5(7):e340, 2016; Bai et al., Current Drug Targets 16:1531-1539, 2015; U.S. Patent Application Publications Nos. 2008/0097091, 2009/0306356, and 2005/0227935; and WO 14/168264), short hairpin RNA (shRNA) (e.g., Jakobsen et al., Mol.
Ther. 17(10): 1743-1753, 2009; Ogawa et al., PLoS One 9(3): e92073, 2014; Ding et al., Bone Joint 94-6(Suppl. 11):44, 2014; and Hernandez-Alejandro et al., J. Surgical Res. 176(2):614-620, 2012), and microRNAs (see, e.g., WO 15/26249). In some embodiments, the inhibitory nucleic acid blocks pre-mRNA splicing of TNFa (e.g., Chiu et al., Mol.
Pharmacol. 71(6):
1640-1645, 2007).
In some embodiments, the inhibitory nucleic acid, e.g., an aptamer (e.g., Orava et al., ACS Chem Biol. 2013; 8(1): 170-178, 2013), can block the binding of a TNFa protein with its receptor (TNFR1 and/or TNFR2).
In some embodiments, the inhibitory nucleic acid can down-regulate the expression of a TNFa-induced downstream mediator (e.g., TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, p38, JNK, IKB-a, or CCL2).
Further teachings of downstream TNFa-induced mediators can be found in, e.g., Schwamborn et al., BMC Genomics 4:46, 2003; and Zhou et al., Oncogene 22: 2034-2044, 2003, incorporated by reference herein. Additional aspects of inhibitory nucleic acids are described in Aagaard et al., Adv. Drug Delivery Rev. 59(2):75-86, 2007, and Burnett et al., Biotechnol. I 6(9):1130-1146, 2011.
In certain embodiments, a therapeutically effective amount of an inhibitory nucleic acid targeting a nucleic acid encoding a TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, INK, c-jun, MEK3/6, p38, PKR, TTP, or MK2 protein can be administered to a subject (e.g., a human subject) in need thereof In some embodiments, the inhibitory nucleic acid can be about 10 nucleotides to about 40 nucleotides (e.g., about 10 to about 30 nucleotides, about 10 to about 25 nucleotides, about 10 to about 20 nucleotides, about 10 to about 15 nucleotides, 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotides, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36 nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, or 40 nucleotides) in length. One skilled in the art will appreciate that inhibitory nucleic acids may comprise at least one modified nucleic acid at either the 5' or 3'end of DNA or RNA.
As is known in the art, the term "thermal melting point (Tm)" refers to the temperature, under defined ionic strength, pH, and inhibitory nucleic acid concentration, at which 50% of the inhibitory nucleic acids complementary to the target sequence hybridize to the target sequence at equilibrium. In some embodiments, an inhibitory nucleic acid can bind specifically to a target nucleic acid under stingent conditions, e.g., those in which the salt concentration is at least about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30 C. for short oligonucleotides (e.g., 10 to 50 nucleotide). Stringent conditions can also be achieved with the addition of destabilizing agents such as formamide.
In some embodiments of any of the inhibitory nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding any one of TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2) with a Tm of greater than 20 C, greater than 22 C, greater than 24 C, greater than 26 C, greater than 28 C, greater than 30 C, greater than 32 C, greater than 34 C, greater than 36 C, greater than 38 C, greater than 40 C, greater than 42 C, greater than 44 C, greater than 46 C, greater than 48 C, greater than 50 C, greater than 52 C, greater than 54 C, greater than 56 C, greater than 58 C, greater than 60 C, greater than 62 C, greater than 64 C, greater than 66 C, greater than 68 C, greater than 70 C, greater than 72 C, greater than 74 C, greater than 76 C, greater than 78 C, or greater than 80 C, e.g., as measured in phosphate buffered saline using a UV
spectrophotometer.
In some embodiments of any of the inhibitor nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding any one of TNFa, TNFR1, TNFR2, TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, NF-KB, CD14, MyD88, IRAK, lipopolysaccharide binding protein (LBP), TRAF6, ras, raf, MEK1/2, ERK1/2, NIK, IKK, IKB, NF-KB, rac, MEK4/7, JNK, c-jun, MEK3/6, p38, PKR, TTP, or MK2) with a Tm of about 20 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, about 30 C, about 28 C, about 26 C, about 24 C, or about 22 C (inclusive); about 22 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, about 30 C, about 28 C, about 26 C, or about 24 C (inclusive); about 24 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, about 30 C, about 28 C, or about 26 C
(inclusive); about 26 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, about 30 C, or about 28 C (inclusive); about 28 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, or about 30 C (inclusive); about 30 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, or about 32 C (inclusive); about 32 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, or about 34 C (inclusive);
about 34 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, or about 36 C (inclusive); about 36 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 .. C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, or about 38 C
(inclusive); about 38 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, or about 40 C (inclusive); about 40 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, or about 42 C (inclusive); about 42 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, or about 44 C (inclusive); about 44 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, or about 46 C (inclusive); about 46 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, or about 48 C (inclusive); about 48 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, or about 50 C
(inclusive); about 50 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, or about 52 C (inclusive); about 52 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, or about 54 C (inclusive); about 54 C
to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, or about 56 C
(inclusive); about 56 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, or about 58 C
(inclusive); about 58 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, or about 60 C (inclusive); about 60 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, or about 62 C (inclusive); about 62 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, or about 64 C

(inclusive); about 64 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, or about 66 C (inclusive); about 66 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, or about 68 C
(inclusive); about 68 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, or about (inclusive); about 70 C to about 80 C, about 78 C, about 76 C, about 74 C, or about 72 C (inclusive); about 72 C to about 80 C, about 78 C, about 76 C, or about (inclusive); about 74 C to about 80 C, about 78 C, or about 76 C
(inclusive); about 76 C
to about 80 C or about 78 C (inclusive); or about 78 C to about 80 C
(inclusive), In some embodiments, the inhibitory nucleic acid can be formulated in a nanoparticle (e.g., a nanoparticle including one or more synthetic polymers, e.g., Patil et al., Pharmaceutical Nanotechnol. 367:195-203, 2009; Yang et al., ACS Appl. Mater.
Interfaces, doi: 10.1021/acsami.6b16556, 2017; Perepelyuk et al., Mol. Ther. Nucleic Acids 6:259-268, 2017). In some embodiments, the nanoparticle can be a mucoadhesive particle (e.g., nanoparticles having a positively-charged exterior surface) (Andersen et al., Methods Mol.
Biol. 555:77-86, 2009). In some embodiments, the nanoparticle can have a neutrally-charged exterior surface.
In some embodiments, the inhibitory nucleic acid can be formulated, e.g., as a liposome (Buyens et al., I Control Release 158(3): 362-370, 2012; Scarabel et al., Expert Op/n. Drug Deliv. 17:1-14, 2017), a micelle (e.g., a mixed micelle) (Tangsangasaksri et al., BioMacromolecules 17:246-255, 2016; Wu et al., Nanotechnology, doi:
10.1088/1361-6528/aa6519, 2017), a microemulsion (WO 11/004395), a nanoemulsion, or a solid lipid nanoparticle (Sahay et al., Nature Biotechnol. 31:653-658, 2013; and Lin et al., Nanomedicine 9(1):105-120, 2014). Additional exemplary structural features of inhibitory nucleic acids and formulations of inhibitory nucleic acids are described in US
2016/0090598.
In some embodiments, a pharmaceutical composition can include a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In some examples, a pharmaceutical composition consists of a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In certain embodiments, the sterile saline is a pharmaceutical grade .. saline. In certain embodiments, a pharmaceutical composition can include one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition includes one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and phosphate-buffered saline (PBS). In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) and sterile phosphate-buffered saline (PBS). In some examples, the sterile saline is a pharmaceutical grade PBS.
In certain embodiments, one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) may be admixed with pharmaceutically acceptable active and/or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions depend on a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.
Pharmaceutical compositions including one or more inhibitory nucleic acids encompass any pharmaceutically acceptable salts, esters, or salts of such esters. Non-limiting examples of pharmaceutical compositions include pharmaceutically acceptable salts of inhibitory nucleic acids. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.
Also provided herein are prodrugs that can include additional nucleosides at one or both ends of an inhibitory nucleic acid which are cleaved by endogenous nucleases within the body, to form the active inhibitory nucleic acid.
Lipid moieties can be used to formulate an inhibitory nucleic acid. In certain such methods, the inhibitory nucleic acid is introduced into preformed liposomes or lipoplexes made of mixtures of cationic lipids and neutral lipids. In certain methods, inhibitory nucleic acid complexes with mono- or poly-cationic lipids are formed without the presence of a neutral lipid. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to a particular cell or tissue in a mammal. In some examples, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to fat tissue in a mammal. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to muscle tissue.
In certain embodiments, pharmaceutical compositions provided herein comprise one or more inhibitory nucleic acid and one or more excipients. In certain such embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose and polyvinylpyrrolidone.
In some examples, a pharmaceutical composition provided herein includes liposomes and emulsions. Liposomes and emulsions can be used to formulate hydrophobic compounds.
In some examples, certain organic solvents such as dimethylsulfoxide are used.
In some examples, a pharmaceutical composition provided herein includes one or more tissue-specific delivery molecules designed to deliver one or more inhibitory nucleic acids to specific tissues or cell types in a mammal. For example, a pharmaceutical composition can include liposomes coated with a tissue-specific antibody.
In some embodiments, a pharmaceutical composition provided herein can include a co-solvent system. Examples of such co-solvent systems include benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. A non-limiting example of such a co-solvent system is the VPD co-solvent system, which is a solution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80 and 65% w/v polyethylene glycol 300. As can be appreciated, other surfactants may be used instead of Polysorbate 80TM; the fraction size of polyethylene glycol may be varied;
other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone;
and other sugars or polysaccharides may substitute for dextrose.
In some examples, a pharmaceutical composition can be formulated for oral administration. In some examples, pharmaceutical compositions are formulated for buccal administration.
In some examples, a pharmaceutical composition is formulated for administration by injection (e.g., intravenous, subcutaneous, intramuscular, etc.). In some of these embodiments, a pharmaceutical composition includes a carrier and is formulated in aqueous solution, such as water or physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. In some examples, other ingredients are included (e.g., ingredients that aid in solubility or serve as preservatives).
In some examples, injectable suspensions are prepared using appropriate liquid carriers, suspending agents, and the like. Some pharmaceutical compositions for injection are formulated in unit dosage form, e.g., in ampoules or in multi-dose containers. Some pharmaceutical compositions for injection are suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, and/or dispersing agents.
Solvents suitable for use in pharmaceutical compositions for injection include, but are not limited to, lipophilic solvents and fatty oils, such as sesame oil, synthetic fatty acid esters, such as ethyl oleate or triglycerides, and liposomes.
Antibodies In some embodiments, the TNFa inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to any one of TNFa, TNFR1, or TNFR2.
In some embodiments, an antibody or antigen-binding fragment of an antibody described herein can bind specifically to TNFa. In some embodiments, an antibody or antigen-binding fragment of an antibody described herein can bind specifically to an TNFa receptor (TNFR1 or TNFR2).
In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc, a VHH domain, a VNAR domain, a (scFv)2, a minibody, or a BiTE. In .. some embodiments, an antibody can be a DVD-Ig, and a dual-affinity re-targeting antibody (DART), a triomab, kih IgG with a common LC, a crossmab, an ortho-Fab IgG, a 2-in-1-IgG, IgG-ScFv, scFv2-Fc, a bi-nanobody, tanden antibody, a DART-Fc, a scFv-HAS-scFv, DNL-Fab3, DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, la-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH
IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody, nanobody-HSA, a diabody, a TandAb, scDiabody, scDiabody-CH3, Diabody-CH3, Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab')2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody, dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HAS, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.
Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab')2 fragment, and a Fab' fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgGl, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgGl, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgAl or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgAl or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).
Non-limiting examples of TNF inhibitors that are antibodies that specifically bind to TNFa are described in Elliott et al., Lancet 1994; 344: 1125-1127, 1994;
Rankin et al., Br.
Rheumatol. 2:334-342, 1995; Butler et al., Eur. Cytokine Network 6(4):225-230, 1994;
Lorenz et al., I Immunol. 156(4):1646-1653, 1996; Hinshaw et al., Circulatory Shock 30(3):279-292, 1990; Wanner et al., Shock 11(6):391-395, 1999; Bongartz et al., AMA
295(19):2275-2285, 2006; Knight et al., Molecular Immunol. 30(16):1443-1453, 1993;
Feldman, Nature Reviews Immunol. 2(5):364-371, 2002; Taylor et al., Nature Reviews Rheumatol. 5(10):578-582, 2009; Garces et al., Annals Rheumatic Dis.
72(12):1947-1955, 2013; Palladino et al., Nature Rev. Drug Discovery 2(9):736-746, 2003;
Sandborn et al., Inflammatory Bowel Diseases 5(2):119-133, 1999; Atzeni et al., Autoimmunity Reviews

12(7):703-708, 2013; Maini et al., Immunol. Rev. 144(1):195-223, 1995; Ordas et al., Clin.
Pharmacol. Therapeutics 91(4):635-646, 2012; Cohen et al., Canadian I
Gastroenterol.
Hepatol. 15(6):376-384, 2001; Feldmann et al., Ann. Rev. Immunol. 19(1):163-196, 2001;
Ben-Horin et al., Autoimmunity Rev. 13(1):24-30, 2014; and U.S. Patent Nos.
6,090,382;
6,258,562; and 6,509,015).
In certain embodiments, the TNFa inhibitor can include or is infliximab (RemicadeTm), CDP571, CDP 870, golimumab (golimumabTM), adalimumab (HumiraTm), or certolizumab pegol (CimziaTm). In certain embodiments, the TNFa inhibitor can be a TNFa inhibitor biosimilar. Examples of approved and late-phase TNFa inhibitor biosimilars include, but are not limited to, infliximab biosimilars such as RemsimaTM and Inflectra (CT-P13) from Celltrion/Pfizer, G5071 from Aprogen, FlixabiTM (5B2) from Samsung Bioepis, PF-06438179 from Pfizer/Sandoz, NI-071 from Nichi-Iko Pharmaceutical Co., and from Amgen; adalimumab biosimilars such as ExemptiaTM (ZRC3197) from Zydus Cadila, India, Solymbic and Amgevita (ABP 501) from Amgen, Imraldi (SB5) from Samsung Bioepis, GP-2017 from Sandoz, Switzerland, ONS-3010 from Oncobiologics/Viropro, U.S.A., M923 from Momenta Pharmaceuticals/Baxalta (Baxter spinoff USA), PF-from Pfizer, BMO-2 or MYL-1401-A from Biocon/Mylan, CHS-1420 from Coherus, FKB327 from Fujifilm/Kyowa Hakko Kirin (Fujifilm Kyowa Kirin Biologics), Cyltezo (BI

695501) from Boehringer Ingelheim, CT-P17 from Celltrion, BAX 923 from Baxalta (now a part of Shire), MSB11022 from Fresenius Kabi (bought from Merck kGaA (Merck Group) in 2017), LBAL from LG Life Sciences/Mochida Pharmaceutical, South Korea/Japan, from Prestige Biopharma, Adfrar from Torrent Pharmaceuticals, India, a biosimilar of adalimumab in development by Adello Biologics, a biosimilar of adalimumab in development by AET Biotech/BioXpress Therapeutics, Germany/Switzerland, a biosimilar of adalimumab from mAbxience, Spain, a biosimilar of adalimumab in development by PlantForm, Canada;
and etanercept biosimilars such as ErelziTM from Sandoz/Novartis, BrenzysTM
(5B4) from Samsung Bioepis, GP2015 from Sandoz, TuNEX from Mycenax, LBEC0101 from LG
Life, and CHS-0214 from Coherus.
In some embodiments, a biosimilar is an antibody or antigen-binding fragment thereof that has a light chain that has the same primary amino acid sequence as compared to a reference antibody (e.g., adalimumab) and a heavy chain that has the same primary amino acid sequence as compared to the reference antibody. In some examples, a biosimilar is an antibody or antigen-binding fragment thereof that has a light chain that includes the same light chain variable domain sequence as a reference antibody (e.g., adalimumab) and a heavy chain that includes the same heavy chain variable domain sequence as a reference antibody.
In some embodiments, a biosimilar can have a similar glycosylation pattern as compared to the reference antibody (e.g., adalimumab). In other embodiments, a biosimilar can have a .. different glycosylation pattern as compared to the reference antibody (e.g., adalimumab).
Changes in the N-linked glycosylation profile of a biosimilar as compared to a reference antibody (e.g., adalimumab) can be detected using 2-anthranilic acid (AA)-derivatization and normal phase liquid chromatography with fluorescence detection, as generally described in Kamoda et al., I Chromatography 1 1133:332-339, 2006.
For example, a biosimilar can have changes in one or more (e.g., two, three, four, five, six, seven, eight, nine, ten, or eleven) of the following types of N-glycosylation as compared to the reference antibody (e.g., adalimumab): neutrally-charged oligosaccharides;
monosialylated fucose-containing oligosaccharides; monosialylated oligosaccharides;
bisialylated fucose-containing oligosaccharide; bisialylated oligosaccharides; triantennary, trisiaylated oligosaccharides of form 1; triantennary, trisialylated oligosaccharides of form 2; mannose-6-phosphate oligosaccharides; monophosphorylated oligosaccharides;
tetrasialylated oligosaccharides; monosialylated and monophosphorylated oligosaccharides; and bis-mannose-6-phosphate oligosaccharides.

In some embodiments, the biosimilar can have a change in one, two, or three of: the percentage of species having one C-terminal lysine, the percentage of species having two C-terminal lysines, and the percentage of species having three C-terminal lysines as compared to the reference antibody (e.g., adalimumab).
In some embodiments, the biosimilar can have a change in the level of one, two, or three of acidic species, neutral species, and basic species in the composition as compared to the reference antibody (e.g., adalimumab).
In some embodiments, the biosimilar can have a change in the level of sulfation as compared to the reference antibody.
In some embodiments, the TNFa inhibitor can be SAR252067 (e.g., a monoclonal antibody that specifically binds to TNFSF14, described in U.S. Patent Application Publication No. 2013/0315913) or MDGN-002 (described in U.S. Patent Application Publication No. 2015/0337046). In some embodiments, the TNFa inhibitor can be PF-06480605, which binds specifically to TNFSF15 (e.g., described in U.S. Patent Application Publication No. 2015/0132311). Additional examples of TNFa inhibitors include (described in Tsianakas et al., Exp. Dermatol. 25:428-433, 2016) and PF-06480605, which binds specifically to TNFSF15 (described in U.S. Patent Application Publication No.
2015/0132311). Further examples of TNFa inhibitors that are antibodies or antigen-binding antibody fragments are described in, e.g., WO 17/158097, EP 3219727, WO
16/156465, and WO 17/167997.
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (KD) of less than 1 x 10-5M (e.g., less than 0.5 x 10-5M, less than 1 x 10' M, less than 0.5 x 10' M, less than 1 x 10-7M, less than 0.5 x 10-7 M, less than 1 x 10-8M, less than 0.5 x 10-8M, less than 1 x 10-9M, less than 0.5 x 10-9M, less than 1 x 10' M, less than 0.5 x 10-1 M, less than 1 x 10-11M, less than 0.5 x 10-11M, or less than 1 x 10-12M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a KD of about lx 10-12 M to about lx 10-5M, about 0.5 x 10-5 M, about lx 10' M, about 0.5 x 10' M, about 1 x 107M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8 M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-1 M, about 0.5 x 10-1 M, about 1 x 10-11M, or about 0.5 x 10-11M (inclusive); about 0.5 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 107M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-1 M, about 0.5 x m or about 1 x 10-11M (inclusive); about 1 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-10M, or about 0.5 x 10-1 M (inclusive); about 0.5 x 10-10M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, or about 1 x 10-1 M
(inclusive);
about 1 x 10-1 M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 107M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9 M, or about 0.5 x 10-9M (inclusive); about 0.5 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, or about 1 x 10-9M (inclusive); about 1 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 107M, about 0.5 x 10-7 M, about 1 x 10-8M, or about 0.5 x 10-8M (inclusive); about 0.5 x 10-8M to about 1 x 10-5 M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, or about 1 x 10-8M (inclusive); about 1 x 10-8M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 107M, or about 0.5 x 10-7M
(inclusive);
about 0.5 x 10-7M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, or about 1 x 10-7M (inclusive); about 1 x 10-7M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, or about 0.5 x 10' M (inclusive); about 0.5 x 10' M to about 1 x 10-5M, about 0.5 x 10-5M, or about 1 x 10' M (inclusive); about 1 x 10-6M to about 1 x 10-5M or about 0.5 x 10-5M (inclusive); or about 0.5 x 10-5M to about 1 x 10-5M
(inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Koff of about lx 10-6 s-1 to about lx 10-3 s-1, about 0.5 x 10-3 s-1, about lx 10' s-1, about 0.5 x 10-4 s-1, about 1 x 10-5 s-1, or about 0.5 x 10-5 s-1 (inclusive); about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10' s-1, about 0.5 x 10' s-1, or about 1 x 10-5 s-1 (inclusive); about 1 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10' s-1, or about 0.5 x 10' s-1 (inclusive); about 0.5 x 10' s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, or about 1 x 10' s-1 (inclusive); about 1 x 10 s-lto about 1 x 10-3 s-1, or about 0.5 x 10-3 s-1 (inclusive); or about 0.5 x 10-5 s-lto about 1 x 10-3 s-1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Kon of about lx 102 M's' to about lx 106 M's', about 0.5 x 106 M's', about 1 x 105M"ls"1, about 0.5 x 105 M's', about 1 x 104 m-1S-1, about 0.5 x 104 M's', about 1 x 103 M's', or about 0.5 x 103 M's' (inclusive); about 0.5 x 103 M"ls"1 to about 1 x 106M"ls-1, about 0.5 x 106 m-1S-1, about 1 x 105M"ls"1, about 0.5 x 105 M's', about 1 x 104 M's', about 0.5 x 104 m-ls-1, or about 1 x 103 M"ls"1 (inclusive); about 1 x 103 M"ls"lto about 1 x 106M"ls"
1, about 0.5 x 106 M's', about 1 x 105M"ls"1, about 0.5 x 105 M's', about 1 x 104 M's', or about 0.5 x 104 m-ls-1 --is-(inclusive); about 0.5 x 104 m lto about 1 x 106M"ls"1, about 0.5 x s, about 1 x 105M"1-s"1-, about 0.5 x 105 M's', or about 1 x 104 M's' (inclusive);
about 1 x M's' to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105M"is-i, or about 0.5 x 105 M"ls"1(inclusive); about 0.5 x 105M"ls"lto about 1 x 106M"ls"1, about 0.5 x s, or about 1 x 105M1s1(inclusive); about 1 x 105M"ls"1 to about 1 x 106M"ls-i, or about 0.5 x 106 m-ls-1 --ls-(inclusive); or about 0.5 x 106 m 'to about 1 x 106M"ls"1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
Fusion Proteins In some embodiments, the TNFa inhibitory agent is a fusion protein (e.g., an extracellular domain of a TNFR fused to a partner peptide, e.g., an Fc region of an immunoglobulin, e.g., human IgG) (see, e.g., Peppel et al., J. Exp. Med.
174(6):1483-1489, 1991; Deeg et al., Leukemia 16(2):162, 2002) or a soluble TNFR (e.g., TNFR1 or TNFR2) that binds specifically to TNFa. In some embodiments, the TNFa inhibitor includes or is etanercept (EnbrelTm) (see, e.g., WO 91/03553 and WO 09/406,476, incorporated by reference herein). In some embodiments, the TNFa inhibitor includes or is r-TBP-I (e.g., Gradstein et al., J. Acquir. Immune Defic. Syndr. 26(2): 111-117, 2001). In some embodiments, the TNFa inhibitor includes or is a soluble TNFa receptor (e.g., Watt et al., J
Leukoc Biol. 66(6):1005-1013, 1999; Tsao et al., Eur Respir I 14(3):490-495, 1999; Kozak et al., Am. J. Physiol. Reg. Integrative Comparative Physiol. 269(1):R23-R29, 1995; Mohler et al., Immunol. 151(3):1548-1561, 1993; Nophar et al., EMBO 9(10):3269, 1990;
Bjornberg et al., Lymphokine Cytokine Res. 13(3):203-211, 1994; Piguet et al., Eur.
Respiratory J. 7(3):515-518, 1994; and Gray et al., Proc. Natl. Acad. Sci.
U.S.A. 87(19):7380-7384, 1990).

Small Molecules In some embodiments, the TNFa inhibitor is a small molecule. In some embodiments, the TNFa inhibitor is C87 (Ma et al., I Biol. Chem. 289(18):12457-66, 2014).
In some embodiments, the small molecule is LMP-420 (e.g., Haraguchi et al., AIDS Res.
Ther. 3:8, 2006). In some embodiments, the small molecule is a tumor necrosis factor-converting enzyme (TACE) inhibitor (e.g., Moss et al., Nature Clinical Practice Rheumatology 4: 300-309, 2008). In some embodiments, the TACE inhibitor is TMI-005 and BMS-561392. Additional examples of small molecule inhibitors are described in, e.g., He et al., Science 310(5750):1022-1025, 2005.
In some examples, the TNFa inhibitor is a small molecule that inhibits the activity of one of TRADD, TRAF2, MEKK1/4, MEKK4/7, JNK, AP-1, ASK1, RIP, MEKK 3/6, MAPK, NIK, IKK, and NF-KB, in a mammalian cell.
In some examples, the TNFa inhibitor is a small molecule that inhibits the activity of one of CD14, MyD88 (see, e.g., Olson et al., Scientific Reports 5:14246, 2015), IRAK
(Chaudhary et al., I Med. Chem. 58(1):96-110, 2015), lipopolysaccharide binding protein (LBP) (see, e.g., U.S. Patent No. 5,705,398), TRAF6 (e.g., 3-[(2,5-Dimethylphenyl)amino]-1-pheny1-2-propen-1-one), ras (e.g., Baker et al., Nature 497:577-578, 2013), raf (e.g., vemurafenib (PLX4032, RG7204), sorafenib tosylate, PLX-4720, dabrafenib (GSK2118436), GDC-0879, RAF265 (CHIR-265), AZ 628, NVP-BHG712, SB590885, ZM 336372, sorafenib, GW5074, TAK-632, CEP-32496, encorafenib (LGX818), CCT196969, LY3009120, R05126766 (CH5126766), PLX7904, and MLN2480), MEK1/2 (e.g., Facciorusso et al., Expert Review Gastroentrol. Hepatol. 9:993-1003, 2015), ERK1/2 (e.g., Mandal et al., Oncogene 35:2547-2561, 2016), NIK (e.g., Mortier et al., Bioorg. Med. Chem.
Lett. 20:4515-4520, 2010), IKK (e.g., Reilly et al., Nature Med. 19:313-321, 2013), I-KB (e.g., Suzuki et al., Expert. Op/n. Invest. Drugs 20:395-405, 2011), NF-KB (e.g., Gupta et al., Biochim. Biophys. Acta 1799(10-12):775-787, 2010), rac (e.g., U.S. Patent No.
9,278,956), MEK4/7, JNK (e.g., AEG 3482, BI 78D3, CEP 1347, c-JUN peptide, IQ 1S, JIP-1 (153-163), 5P600125, SU 3327, and TCS JNK6o), c-jun (e.g., AEG 3482, BI 78D3, CEP 1347, c-JUN
peptide, IQ 1S, JIP-1 (153-163), 5P600125, SU 3327, and TCS JNK6o), MEK3/6 (e.g., Akinleye et al., I Hematol. Oncol. 6:27, 2013), p38 (e.g., AL 8697, AMG 548, BIRB 796, CMPD-1, DBM 1285 dihydrochloride, EO 1428, JX 401, ML 3403, Org 48762-0, PH
797804, RWJ 67657, SB 202190, SB 203580, SB 239063, SB 706504, SCIO 469, SKF
86002, SX 011, TA 01, TA 02, TAK 715, VX 702, and VX 745), PKR (e.g., 2-aminopurine or CAS 608512-97-6), TTP (e.g., CAS 329907-28-0), and MK2 (PF 3644022 and PHA
767491).
IL-6 Receptor Inhibitors The term "IL-6 receptor inhibitor" refers to an agent which decreases IL-6 receptor expression and/or the ability of IL-6 to bind to an IL-6 receptor. In some embodiments, the IL-6 receptor inhibitor targets the IL-6 receptor 13-subunit, glycoprotein 130 (sIL6gp130). In other embodiments, the IL-6 receptor inhibitor targets the IL-6 receptor subunit (IL6R). In other embodiments, the IL-6 receptor inhibitor targets the complex consisting of both the IL-6 receptor subunit (IL6R) and the IL-6 receptor 13-subunit, glycoprotein 130 (sIL6gp130). In some embodiments, the IL-6 receptor inhibitor targets IL-6.
In some embodiments, an IL-6 receptor inhibitor is an inhibitory nucleic acid, an antibody or an antigen-binding fragment thereof, a fusion protein, a IL-6 receptor antagonist, or a small molecule. In some embodiments, the inhibitory nucleic acid is a small interfering RNA, an antisense nucleic acid, an aptamer, or a microRNA. Exemplary IL-6 receptor inhibitors are described herein. Additional examples of IL-6 receptor inhibitors are known in the art.
Exemplary aspects of different inhibitory nucleic acids are described below.
Any of the examples of inhibitory nucleic acids that can decrease expression of an IL6R, sIL6gp130, or IL-6 mRNA. Inhibitory nucleic acids that can decrease the expression of IL6R, sIL6gp130, or IL-6 mRNA in a mammalian cell include antisense nucleic acid molecules, i.e., nucleic acid molecules whose nucleotide sequence is complementary to all or part of an IL6R, sIL6gp130, or IL-6 mRNA (e.g., complementary to all or a part of any one of SEQ ID
NOs: 50-55).
Human IL6R mRNA Variant 1 (SEQ ID NO: 50) 1 ggcggtcccc tgttctcccc gctcaggtgc ggcgctgtgg caggaagcca ccccctcggt 61 cggccggtgc gcggggctgt tgcgccatcc gctccggctt tcgtaaccgc accctgggac 121 ggcccagaga cgctccagcg cgagttcctc aaatgttttc ctgcgttgcc aggaccgtcc 181 gccgctctga gtcatgtgcg agtgggaagt cgcactgaca ctgagccggg ccagagggag 241 aggagccgag cgcggcgcgg ggccgaggga ctcgcagtgt gtgtagagag ccgggctcct 301 gcggatgggg gctgcccccg gggcctgagc ccgcctgccc gcccaccgcc ccgccccgcc 361 cctgccaccc ctgccgcccg gttcccatta gcctgtccgc ctctgcggga ccatggagtg 421 gtagccgagg aggaagcatg ctggccgtcg gctgcgcgct gctggctgcc ctgctggccg 481 cgccgggagc ggcgctggcc ccaaggcgct gccctgcgca ggaggtggcg agaggcgtgc 541 tgaccagtct gccaggagac agcgtgactc tgacctgccc gggggtagag ccggaagaca 601 atgccactgt tcactgggtg ctcaggaagc cggctgcagg ctcccacccc agcagatggg 661 ctggcatggg aaggaggctg ctgctgaggt cggtgcagct ccacgactct ggaaactatt 721 catgctaccg ggccggccgc ccagctggga ctgtgcactt gctggtggat gttccccccg 781 aggagcccca gctctcctgc ttccggaaga gccccctcag caatgttgtt tgtgagtggg 841 gtcctcggag caccccatcc ctgacgacaa aggctgtgct cttggtgagg aagtttcaga 901 acagtccggc cgaagacttc caggagccgt gccagtattc ccaggagtcc cagaagttct 961 cctgccagtt agcagtcccg gagggagaca gctctttcta catagtgtcc atgtgcgtcg 1021 ccagtagtgt cgggagcaag ttcagcaaaa ctcaaacctt tcagggttgt ggaatcttgc 1081 agcctgatcc gcctgccaac atcacagtca ctgccgtggc cagaaacccc cgctggctca 1141 gtgtcacctg gcaagacccc cactcctgga actcatcttt ctacagacta cggtttgagc 1201 tcagatatcg ggctgaacgg tcaaagacat tcacaacatg gatggtcaag gacctccagc 1261 atcactgtgt catccacgac gcctggagcg gcctgaggca cgtggtgcag cttcgtgccc 1321 aggaggagtt cgggcaaggc gagtggagcg agtggagccc ggaggccatg ggcacgcctt 1381 ggacagaatc caggagtcct ccagctgaga acgaggtgtc cacccccatg caggcactta 1441 ctactaataa agacgatgat aatattctct tcagagattc tgcaaatgcg acaagcctcc 1501 cagtgcaaga ttcttcttca gtaccactgc ccacattcct ggttgctgga gggagcctgg 1561 ccttcggaac gctcctctgc attgccattg ttctgaggtt caagaagacg tggaagctgc 1621 gggctctgaa ggaaggcaag acaagcatgc atccgccgta ctctttgggg cagctggtcc 1681 cggagaggcc tcgacccacc ccagtgcttg ttcctctcat ctccccaccg gtgtccccca 1741 gcagcctggg gtctgacaat acctcgagcc acaaccgacc agatgccagg gacccacgga 1801 gcccttatga catcagcaat acagactact tcttccccag atagctggct gggtggcacc 1861 agcagcctgg accctgtgga tgataaaaca caaacgggct cagcaaaaga tgcttctcac 1921 tgccatgcca gcttatctca ggggtgtgcg gcctttggct tcacggaaga gccttgcgga 1981 aggttctacg ccaggggaaa atcagcctgc tccagctgtt cagctggttg aggtttcaaa 2041 cctccctttc caaatgccca gcttaaaggg gctagagtga acttgggcca ctgtgaagag 2101 aaccatatca agactctttg gacactcaca cggacactca aaagctgggc aggttggtgg 2161 gggcctcggt gtggagaagc ggctggcagc ccacccctca acacctctgc acaagctgca 2221 ccctcaggca ggtgggatgg atttccagcc aaagcctcct ccagccgcca tgctcctggc 2281 ccactgcatc gtttcatctt ccaactcaaa ctcttaaaac ccaagtgcct tagcaaattc 2341 tgtttttcta ggcctgggga cggcttttac ttaaaccgcc aaggctgggg gaagaagctc 2401 tctcctccct ttcttcccta cagttgaaaa acagctgagg gtgagtgggt gaataataca 2461 gtatctcagg gcctggtcgt tttcaacaga attataatta gttcctcatt agcattttgc 2521 taaatgtgaa tgatgatcct aggcatttgc tgaatacaga ggcaactgca ttggctttgg 2581 gttgcaggac ctcaggtgag aagcagagga aggagaggag aggggcacag ggtctctacc 2641 atcccctgta gagtgggagc tgagtggggg atcacagcct ctgaaaacca atgttctctc 2701 ttctccacct cccacaaagg agagctagca gcagggaggg cttctgccat ttctgagatc 2761 aaaacggttt tactgcagct ttgtttgttg tcagctgaac ctgggtaact agggaagata 2821 atattaagga agacaatgtg aaaagaaaaa tgagcctggc aagaatgtgt ttaaacttgg 2881 tttttaaaaa actgctgact gttttctctt gagagggtgg aatatccaat attcgctgtg 2941 tcagcataga agtaacttac ttaggtgtgg gggaagcacc ataactttgt ttagcccaaa 3001 accaagtcaa gtgaaaaagg aggaagagaa aaaatatttt cctgccaggc atggtggccc 3061 acgcacttcg ggaggtcgag gcaggaggat cacttgagtc cagaagtttg agatcagcct 3121 gggcaatgtg ataaaacccc atctctacaa aaagcataaa aattagccaa gtgtggtaga 3181 gtgtgcctga agtcccagat acttgggggg ctgaggtggg aggatctctt gagcctggga 3241 ggtcaaggct gcagtgagcc gagattgcac cactgcactc cagcctgggt gacagagcaa 3301 gtgagaccct gtctcaaaaa aagaaaaaga aaaagaaaaa atattttccc tattagagaa 3361 gagattgtgg tttcattctg tattttgttt ttgtcttaaa aagtggaaaa atagcctgcc 3421 tcttctctac tctagggaaa aaccagcgtg tgactactcc cccaggtggt tatggagagg 3481 gtgtccggtc cctgtcccag tgccgagaag gaagcctccc acgactgccc ggcagggtcc 3541 tagaaattcc ccaccctgaa agccctgagc tttctgctat caaagaggtt ttaaaaaaat 3601 cccatttaaa aaaaatccct tacctcggtg ccttcctctt tttatttagt tccttgagtt 3661 gattcagctc tgcaagaatt gaagcaggac taaatgtcta gttgtaacac catgattaac 3721 cacttcagct gacttttctg tccgagcttt gaaaattcag tggtgttagt ggttacccag 3781 ttagctctca agttatcagg gtattccaga gtggggatat gatttaaatc agccgtgtaa 3841 ccatggaccc aatatttacc agaccacaaa acttttctaa tactctaccc tcttagaaaa 3901 accaccacca tcaccagaca ggtgcgaaag gatgaaagtg accatgtttt gtttacggtt 3961 ttccaggttt aagctgttac tgtcttcagt aagccgtgat tttcattgct gggcttgtct 4021 gtagatttta gaccctattg ctgcttgagg caactcatct taggttggca aaaaggcagg 4081 atggccgggc gcggtggctc acgcctgtaa tcctagcact ttgggaggcc aaggtgggag 4141 gattgcttga gctcaggagt ttgagaccaa cctgggtaac atagtgagac accatctcta 4201 ttatgaacaa taacagttaa gaaaaaaaaa ggcaggcagg cggttatggt ggttccctcc 4261 catcccacca cataaagttt ctgagacttg agaacagcaa aatgctgtta aagggaaata 4321 ttaagaatga gaatctgcag taagggtgat tctgtgccca cagttcttca attctttata 4381 ccgttttacc cacatgtggt gttaccaaag ccgggcagaa ccatgctagc ggaagatgtg 4441 aaatccagat agctcattat tgccaagagc taggcagctt tgatctccaa attgttattg 4501 ctttcatttt tattgtaatg gaattgcttt gttttgtttt tttgtttttg tattgaagag 4561 ggttgttttc cctttatttt tcataagcta atgtaaatga agaaaaaatg tcttctctgg 4621 gctgtaggcc tggctcagcg tacacaggta tacatcctaa gctctctatg ttctctaatc 4681 tgtggtgact gaacatgtgt ctcaatgcac ggggcatttc tacctgtgtt tctgcagcac 4741 ccccactgcc ttgagtcccc agcagtgctg ttatttgcct aacacctgta gccatctgcc 4801 acgcagccag acgtgaaacg ctgagacaga gaccatttag gttaaatacg acagcttatc 4861 ctgctgggtg gggaaagtaa aaaatatgct ggttcaaggc ctaaagtaaa atgatcaata 4921 atgtttgtag cattaatgaa atattttcaa gaaatgtgtc caggggtagc actggctatg 4981 ttgacgaggc ctttggtaac tcagagagct cttggccctg atggggactt gcccttacgc 5041 tttctttatc aggctctgag ttcacacgga gcctctggca cttccctgct gtcttgggag 5101 aaaggaaact ggttgccgcg gcaggttgtg gaatctgttg ctggaaccag gctggaagcc 5161 cacctggtag tgaacagggc ccagtggggc aggctgggca tgttgtggtc tatgggtttg 5221 tttcctggag aatgttcagg aatgtcttcc cagctgcttt ggtgctgagc tctattatct 5281 cacagcacgt ccagaaggct aacccaggtg gggaggatgc tgacaccagc tccaggtgga 5341 gttggtggtc ttaatttgga gatgcagggg caacctgtga ccctttgagg caagagccct 5401 gcacccagct gtcccgtgca gccgtgggca ggggctgcac acggaggggc aggcgggcca 5461 gttcagggtc cgtgccaggc cctcctcagt gccctgtgaa ggcctcctgt cctccgtgcg 5521 gctgggcacc agcaccaggg agtttctatg gcaaccttag tgattattaa ggaacactgt 5581 cagttttatg aacatatgct caaatgaaat tctactttag gaggaaagga ttggaacagc 5641 atgtcacaag gctgttaatt aacagagaga ccttattgga tggagatcac atctgttaaa 5701 tagaatacct caactctacg ttgttttctt ggagataaat aatagtttca agtttttgtt 5761 tgtttgtttt acctaattac ctgaaagcaa ataccaaagg ctgatgtctg tatatggggc 5821 aaagggtcag tatatttttc agtgtttttt tttctaccag ctattttgca tttaaagtga 5881 acattgtgtt tggaataaat actcttaaaa aataaaaaaa aaaaaaaa Human IL6R mRNA Variant 2 (SEQ ID NO: 51) 1 ggcggtcccc tgttctcccc gctcaggtgc ggcgctgtgg caggaagcca ccccctcggt 61 cggccggtgc gcggggctgt tgcgccatcc gctccggctt tcgtaaccgc accctgggac 121 ggcccagaga cgctccagcg cgagttcctc aaatgttttc ctgcgttgcc aggaccgtcc 181 gccgctctga gtcatgtgcg agtgggaagt cgcactgaca ctgagccggg ccagagggag 241 aggagccgag cgcggcgcgg ggccgaggga ctcgcagtgt gtgtagagag ccgggctcct 301 gcggatgggg gctgcccccg gggcctgagc ccgcctgccc gcccaccgcc ccgccccgcc 361 cctgccaccc ctgccgcccg gttcccatta gcctgtccgc ctctgcggga ccatggagtg 421 gtagccgagg aggaagcatg ctggccgtcg gctgcgcgct gctggctgcc ctgctggccg 481 cgccgggagc ggcgctggcc ccaaggcgct gccctgcgca ggaggtggcg agaggcgtgc 541 tgaccagtct gccaggagac agcgtgactc tgacctgccc gggggtagag ccggaagaca 601 atgccactgt tcactgggtg ctcaggaagc cggctgcagg ctcccacccc agcagatggg 661 ctggcatggg aaggaggctg ctgctgaggt cggtgcagct ccacgactct ggaaactatt 721 catgctaccg ggccggccgc ccagctggga ctgtgcactt gctggtggat gttccccccg 781 aggagcccca gctctcctgc ttccggaaga gccccctcag caatgttgtt tgtgagtggg 841 gtcctcggag caccccatcc ctgacgacaa aggctgtgct cttggtgagg aagtttcaga 901 acagtccggc cgaagacttc caggagccgt gccagtattc ccaggagtcc cagaagttct 961 cctgccagtt agcagtcccg gagggagaca gctctttcta catagtgtcc atgtgcgtcg 1021 ccagtagtgt cgggagcaag ttcagcaaaa ctcaaacctt tcagggttgt ggaatcttgc 1081 agcctgatcc gcctgccaac atcacagtca ctgccgtggc cagaaacccc cgctggctca 1141 gtgtcacctg gcaagacccc cactcctgga actcatcttt ctacagacta cggtttgagc 1201 tcagatatcg ggctgaacgg tcaaagacat tcacaacatg gatggtcaag gacctccagc 1261 atcactgtgt catccacgac gcctggagcg gcctgaggca cgtggtgcag cttcgtgccc 1321 aggaggagtt cgggcaaggc gagtggagcg agtggagccc ggaggccatg ggcacgcctt 1381 ggacagaatc caggagtcct ccagctgaga acgaggtgtc cacccccatg caggcactta 1441 ctactaataa agacgatgat aatattctct tcagagattc tgcaaatgcg acaagcctcc 1501 caggttcaag aagacgtgga agctgcgggc tctgaaggaa ggcaagacaa gcatgcatcc 1561 gccgtactct ttggggcagc tggtcccgga gaggcctcga cccaccccag tgcttgttcc 1621 tctcatctcc ccaccggtgt cccccagcag cctggggtct gacaatacct cgagccacaa 1681 ccgaccagat gccagggacc cacggagccc ttatgacatc agcaatacag actacttctt 1741 ccccagatag ctggctgggt ggcaccagca gcctggaccc tgtggatgat aaaacacaaa 1801 cgggctcagc aaaagatgct tctcactgcc atgccagctt atctcagggg tgtgcggcct 1861 ttggcttcac ggaagagcct tgcggaaggt tctacgccag gggaaaatca gcctgctcca 1921 gctgttcagc tggttgaggt ttcaaacctc cctttccaaa tgcccagctt aaaggggcta 1981 gagtgaactt gggccactgt gaagagaacc atatcaagac tctttggaca ctcacacgga 2041 cactcaaaag ctgggcaggt tggtgggggc ctcggtgtgg agaagcggct ggcagcccac 2101 ccctcaacac ctctgcacaa gctgcaccct caggcaggtg ggatggattt ccagccaaag 2161 cctcctccag ccgccatgct cctggcccac tgcatcgttt catcttccaa ctcaaactct 2221 taaaacccaa gtgccttagc aaattctgtt tttctaggcc tggggacggc ttttacttaa 2281 accgccaagg ctgggggaag aagctctctc ctccctttct tccctacagt tgaaaaacag 2341 ctgagggtga gtgggtgaat aatacagtat ctcagggcct ggtcgttttc aacagaatta 2401 taattagttc ctcattagca ttttgctaaa tgtgaatgat gatcctaggc atttgctgaa 2461 tacagaggca actgcattgg ctttgggttg caggacctca ggtgagaagc agaggaagga 2521 gaggagaggg gcacagggtc tctaccatcc cctgtagagt gggagctgag tgggggatca 2581 cagcctctga aaaccaatgt tctctcttct ccacctccca caaaggagag ctagcagcag 2641 ggagggcttc tgccatttct gagatcaaaa cggttttact gcagctttgt ttgttgtcag 2701 ctgaacctgg gtaactaggg aagataatat taaggaagac aatgtgaaaa gaaaaatgag 2761 cctggcaaga atgtgtttaa acttggtttt taaaaaactg ctgactgttt tctcttgaga 2821 gggtggaata tccaatattc gctgtgtcag catagaagta acttacttag gtgtggggga 2881 agcaccataa ctttgtttag cccaaaacca agtcaagtga aaaaggagga agagaaaaaa 2941 tattttcctg ccaggcatgg tggcccacgc acttcgggag gtcgaggcag gaggatcact 3001 tgagtccaga agtttgagat cagcctgggc aatgtgataa aaccccatct ctacaaaaag 3061 cataaaaatt agccaagtgt ggtagagtgt gcctgaagtc ccagatactt ggggggctga 3121 ggtgggagga tctcttgagc ctgggaggtc aaggctgcag tgagccgaga ttgcaccact 3181 gcactccagc ctgggtgaca gagcaagtga gaccctgtct caaaaaaaga aaaagaaaaa 3241 gaaaaaatat tttccctatt agagaagaga ttgtggtttc attctgtatt ttgtttttgt 3301 cttaaaaagt ggaaaaatag cctgcctctt ctctactcta gggaaaaacc agcgtgtgac 3361 tactccccca ggtggttatg gagagggtgt ccggtccctg tcccagtgcc gagaaggaag 3421 cctcccacga ctgcccggca gggtcctaga aattccccac cctgaaagcc ctgagctttc 3481 tgctatcaaa gaggttttaa aaaaatccca tttaaaaaaa atcccttacc tcggtgcctt 3541 cctcttttta tttagttcct tgagttgatt cagctctgca agaattgaag caggactaaa 3601 tgtctagttg taacaccatg attaaccact tcagctgact tttctgtccg agctttgaaa 3661 attcagtggt gttagtggtt acccagttag ctctcaagtt atcagggtat tccagagtgg 3721 ggatatgatt taaatcagcc gtgtaaccat ggacccaata tttaccagac cacaaaactt 3781 ttctaatact ctaccctctt agaaaaacca ccaccatcac cagacaggtg cgaaaggatg 3841 aaagtgacca tgttttgttt acggttttcc aggtttaagc tgttactgtc ttcagtaagc 3901 cgtgattttc attgctgggc ttgtctgtag attttagacc ctattgctgc ttgaggcaac 3961 tcatcttagg ttggcaaaaa ggcaggatgg ccgggcgcgg tggctcacgc ctgtaatcct 4021 agcactttgg gaggccaagg tgggaggatt gcttgagctc aggagtttga gaccaacctg 4081 ggtaacatag tgagacacca tctctattat gaacaataac agttaagaaa aaaaaaggca 4141 ggcaggcggt tatggtggtt ccctcccatc ccaccacata aagtttctga gacttgagaa 4201 cagcaaaatg ctgttaaagg gaaatattaa gaatgagaat ctgcagtaag ggtgattctg 4261 tgcccacagt tcttcaattc tttataccgt tttacccaca tgtggtgtta ccaaagccgg 4321 gcagaaccat gctagcggaa gatgtgaaat ccagatagct cattattgcc aagagctagg 4381 cagctttgat ctccaaattg ttattgcttt catttttatt gtaatggaat tgctttgttt 4441 tgtttttttg tttttgtatt gaagagggtt gttttccctt tatttttcat aagctaatgt 4501 aaatgaagaa aaaatgtctt ctctgggctg taggcctggc tcagcgtaca caggtataca 4561 tcctaagctc tctatgttct ctaatctgtg gtgactgaac atgtgtctca atgcacgggg 4621 catttctacc tgtgtttctg cagcaccccc actgccttga gtccccagca gtgctgttat 4681 ttgcctaaca cctgtagcca tctgccacgc agccagacgt gaaacgctga gacagagacc 4741 atttaggtta aatacgacag cttatcctgc tgggtgggga aagtaaaaaa tatgctggtt 4801 caaggcctaa agtaaaatga tcaataatgt ttgtagcatt aatgaaatat tttcaagaaa 4861 tgtgtccagg ggtagcactg gctatgttga cgaggccttt ggtaactcag agagctcttg 4921 gccctgatgg ggacttgccc ttacgctttc tttatcaggc tctgagttca cacggagcct 4981 ctggcacttc cctgctgtct tgggagaaag gaaactggtt gccgcggcag gttgtggaat 5041 ctgttgctgg aaccaggctg gaagcccacc tggtagtgaa cagggcccag tggggcaggc 5101 tgggcatgtt gtggtctatg ggtttgtttc ctggagaatg ttcaggaatg tcttcccagc 5161 tgctttggtg ctgagctcta ttatctcaca gcacgtccag aaggctaacc caggtgggga 5221 ggatgctgac accagctcca ggtggagttg gtggtcttaa tttggagatg caggggcaac 5281 ctgtgaccct ttgaggcaag agccctgcac ccagctgtcc cgtgcagccg tgggcagggg 5341 ctgcacacgg aggggcaggc gggccagttc agggtccgtg ccaggccctc ctcagtgccc 5401 tgtgaaggcc tcctgtcctc cgtgcggctg ggcaccagca ccagggagtt tctatggcaa 5461 ccttagtgat tattaaggaa cactgtcagt tttatgaaca tatgctcaaa tgaaattcta 5521 ctttaggagg aaaggattgg aacagcatgt cacaaggctg ttaattaaca gagagacctt 5581 attggatgga gatcacatct gttaaataga atacctcaac tctacgttgt tttcttggag 5641 ataaataata gtttcaagtt tttgtttgtt tgttttacct aattacctga aagcaaatac 5701 caaaggctga tgtctgtata tggggcaaag ggtcagtata tttttcagtg tttttttttc 5761 taccagctat tttgcattta aagtgaacat tgtgtttgga ataaatactc ttaaaaaata 5821 aaaaaaaaaa aaaa Human IL6R mRNA Variant 3 (SEQ ID NO: 52) 1 ggcggtcccc tgttctcccc gctcaggtgc ggcgctgtgg caggaagcca ccccctcggt 61 cggccggtgc gcggggctgt tgcgccatcc gctccggctt tcgtaaccgc accctgggac 121 ggcccagaga cgctccagcg cgagttcctc aaatgttttc ctgcgttgcc aggaccgtcc 181 gccgctctga gtcatgtgcg agtgggaagt cgcactgaca ctgagccggg ccagagggag 241 aggagccgag cgcggcgcgg ggccgaggga ctcgcagtgt gtgtagagag ccgggctcct 301 gcggatgggg gctgcccccg gggcctgagc ccgcctgccc gcccaccgcc ccgccccgcc 361 cctgccaccc ctgccgcccg gttcccatta gcctgtccgc ctctgcggga ccatggagtg 421 gtagccgagg aggaagcatg ctggccgtcg gctgcgcgct gctggctgcc ctgctggccg 481 cgccgggagc ggcgctggcc ccaaggcgct gccctgcgca ggaggtggcg agaggcgtgc 541 tgaccagtct gccaggagac agcgtgactc tgacctgccc gggggtagag ccggaagaca 601 atgccactgt tcactgggtg ctcaggaagc cggctgcagg ctcccacccc agcagatggg 661 ctggcatggg aaggaggctg ctgctgaggt cggtgcagct ccacgactct ggaaactatt 721 catgctaccg ggccggccgc ccagctggga ctgtgcactt gctggtggat gttccccccg 781 aggagcccca gctctcctgc ttccggaaga gccccctcag caatgttgtt tgtgagtggg 841 gtcctcggag caccccatcc ctgacgacaa aggctgtgct cttggtgagg aagtttcaga 901 acagtccggc cgaagacttc caggagccgt gccagtattc ccaggagtcc cagaagttct 961 cctgccagtt agcagtcccg gagggagaca gctctttcta catagtgtcc atgtgcgtcg 1021 ccagtagtgt cgggagcaag ttcagcaaaa ctcaaacctt tcagggttgt ggaatcttgc 1081 agcctgatcc gcctgccaac atcacagtca ctgccgtggc cagaaacccc cgctggctca 1141 gtgtcacctg gcaagacccc cactcctgga actcatcttt ctacagacta cggtttgagc 1201 tcagatatcg ggctgaacgg tcaaagacat tcacaacatg gatggtcaag gacctccagc 1261 atcactgtgt catccacgac gcctggagcg gcctgaggca cgtggtgcag cttcgtgccc 1321 aggaggagtt cgggcaaggc gagtggagcg agtggagccc ggaggccatg ggcacgcctt 1381 ggacagacag gctttctcct cgttgcccag gatggagtac agcagtgcaa tcacagctca 1441 cggcaacttc tgcctcctgg gttcaagcaa tcctcccgcc tcagcctcct aagtagctgg 1501 gaccacaggc gtgtgccaca atgctaattt tttaaaaatg ttttgtagag acagggtttc 1561 accatgctgc ccaggctggt ctcgaactcc tggcctcaag tgatccacca gcctcagact 1621 cccaaagtgc tgggattact ggtgtgagcc actgcacctg actaaacttt aaatttlitt 1681 ttttagacgg aatctcgctc tgttgcccag gctggagtgc agtggcatga tattggctca 1741 ctgcaagctc tgcctcttgg gttcacgcta ttctcctgcc tcagcctcct gagtagctgg 1801 gactacaggt gcacaccacc acgcccggct aattattt tttttttagt agagacgggg 1861 tttcactgtg ttggccaggc tggtcttgaa ctcctgacct cgtgatccac ccgcctcgcc 1921 ctcccaaaat gctgggatta caggtgtgag ccaccgcgcc tggcctaaac ttttaaaatt 1981 ttaatcaaat taatacatgc acatggcaaa gaagtaataa acagcttata acactgaaaa 2041 aaaaaaaaaa aaaaaaaa Human IL-6 receptor I3-subunit, glycoprotein 130 (sIL6gp130) (SEQ ID NO: 53) 1 gagcagccaa aaggcccgcg gagtcgcgct gggccgcccc ggcgcagctg aaccgggggc 61 cgcgcctgcc aggccgacgg gtctggccca gcctggcgcc aaggggttcg tgcgctgtgg 121 agacgcggag ggtcgaggcg gcgcggcctg agtgaaaccc aatggaaaaa gcatgacatt 181 tagaagtaga agacttagct tcaaatccct actccttcac ttactaattt tgtgatttgg 241 aaatatccgc gcaagatgtt gacgttgcag acttgggtag tgcaagcctt gtttattttc 301 ctcaccactg aatctacagg tgaacttcta gatccatgtg gttatatcag tcctgaatct 361 ccagttgtac aacttcattc taatttcact gcagtttgtg tgctaaagga aaaatgtatg 421 gattattttc atgtaaatgc taattacatt gtctggaaaa caaaccattt tactattcct 481 aaggagcaat atactatcat aaacagaaca gcatccagtg tcacctttac agatatagct 541 tcattaaata ttcagctcac ttgcaacatt cttacattcg gacagcttga acagaatgtt 601 tatggaatca caataatttc aggcttgcct ccagaaaaac ctaaaaattt gagttgcatt 661 gtgaacgagg ggaagaaaat gaggtgtgag tgggatggtg gaagggaaac acacttggag 721 acaaacttca ctttaaaatc tgaatgggca acacacaagt ttgctgattg caaagcaaaa 781 cgtgacaccc ccacctcatg cactgttgat tattctactg tgtattttgt caacattgaa 841 gtctgggtag aagcagagaa tgcccttggg aaggttacat cagatcatat caattttgat 901 cctgtatata aagtgaagcc caatccgcca cataatttat cagtgatcaa ctcagaggaa 961 ctgtctagta tcttaaaatt gacatggacc aacccaagta ttaagagtgt tataatacta 1021 aaatataaca ttcaatatag gaccaaagat gcctcaactt ggagccagat tcctcctgaa 1081 gacacagcat ccacccgatc ttcattcact gtccaagacc ttaaaccttt tacagaatat 1141 gtgtttagga ttcgctgtat gaaggaagat ggtaagggat actggagtga ctggagtgaa 1201 gaagcaagtg ggatcaccta tgaagataga ccatctaaag caccaagttt ctggtataaa 1261 atagatccat cccatactca aggctacaga actgtacaac tcgtgtggaa gacattgcct 1321 ccttttgaag ccaatggaaa aatcttggat tatgaagtga ctctcacaag atggaaatca 1381 catttacaaa attacacagt taatgccaca aaactgacag taaatctcac aaatgatcgc 1441 tatctagcaa ccctaacagt aagaaatctt gttggcaaat cagatgcagc tgttttaact 1501 atccctgcct gtgactttca agctactcac cctgtaatgg atcttaaagc attccccaaa 1561 gataacatgc tttgggtgga atggactact ccaagggaat ctgtaaagaa atatatactt 1621 gagtggtgtg tgttatcaga taaagcaccc tgtatcacag actggcaaca agaagatggt 1681 accgtgcatc gcacctattt aagagggaac ttagcagaga gcaaatgcta tttgataaca 1741 gttactccag tatatgctga tggaccagga agccctgaat ccataaaggc ataccttaaa 1801 caagctccac cttccaaagg acctactgtt cggacaaaaa aagtagggaa aaacgaagct 1861 gtcttagagt gggaccaact tcctgttgat gttcagaatg gatttatcag aaattatact 1921 atattttata gaaccatcat tggaaatgaa actgctgtga atgtggattc ttcccacaca 1981 gaatatacat tgtcctcttt gactagtgac acattgtaca tggtacgaat ggcagcatac 2041 acagatgaag gtgggaagga tggtccagaa ttcactttta ctaccccaaa gtttgctcaa 2101 ggagaaattg aagccatagt cgtgcctgtt tgcttagcat tcctattgac aactcttctg 2161 ggagtgctgt tctgctttaa taagcgagac ctaattaaaa aacacatctg gcctaatgtt 2221 ccagatcctt caaagagtca tattgcccag tggtcacctc acactcctcc aaggcacaat 2281 tttaattcaa aagatcaaat gtattcagat ggcaatttca ctgatgtaag tgttgtggaa 2341 atagaagcaa atgacaaaaa gccttttcca gaagatctga aatcattgga cctgttcaaa 2401 aaggaaaaaa ttaatactga aggacacagc agtggtattg gggggtcttc atgcatgtca 2461 tcttctaggc caagcatttc tagcagtgat gaaaatgaat cttcacaaaa cacttcgagc 2521 actgtccagt attctaccgt ggtacacagt ggctacagac accaagttcc gtcagtccaa 2581 gtcttctcaa gatccgagtc tacccagccc ttgttagatt cagaggagcg gccagaagat 2641 ctacaattag tagatcatgt agatggcggt gatggtattt tgcccaggca acagtacttc 2701 aaacagaact gcagtcagca tgaatccagt ccagatattt cacattttga aaggtcaaag 2761 caagtttcat cagtcaatga ggaagatttt gttagactta aacagcagat ttcagatcat 2821 atttcacaat cctgtggatc tgggcaaatg aaaatgtttc aggaagtttc tgcagcagat 2881 gcttttggtc caggtactga gggacaagta gaaagatttg aaacagttgg catggaggct 2941 gcgactgatg aaggcatgcc taaaagttac ttaccacaga ctgtacggca aggcggctac 3001 atgcctcagt gaaggactag tagttcctgc tacaacttca gcagtaccta taaagtaaag 3061 ctaaaatgat tttatctgtg aattc Human IL-6 mRNA Transcript 1 (SEQ ID NO: 54) 1 gtctcaatat tagagtctca acccccaata aatataggac tggagatgtc tgaggctcat 61 tctgccctcg agcccaccgg gaacgaaaga gaagctctat ctcccctcca ggagcccagc 121 tatgaactcc ttctccacaa gcgccttcgg tccagttgcc ttctccctgg ggctgctcct 181 ggtgttgcct gctgccttcc ctgccccagt acccccagga gaagattcca aagatgtagc 241 cgccccacac agacagccac tcacctcttc agaacgaatt gacaaacaaa ttcggtacat 301 cctcgacggc atctcagccc tgagaaagga gacatgtaac aagagtaaca tgtgtgaaag 361 cagcaaagag gcactggcag aaaacaacct gaaccttcca aagatggctg aaaaagatgg 421 atgcttccaa tctggattca atgaggagac ttgcctggtg aaaatcatca ctggtctttt 481 ggagtttgag gtatacctag agtacctcca gaacagattt gagagtagtg aggaacaagc 541 cagagctgtg cagatgagta caaaagtcct gatccagttc ctgcagaaaa aggcaaagaa 601 tctagatgca ataaccaccc ctgacccaac cacaaatgcc agcctgctga cgaagctgca 661 ggcacagaac cagtggctgc aggacatgac aactcatctc attctgcgca gctttaagga 721 gttcctgcag tccagcctga gggctcttcg gcaaatgtag catgggcacc tcagattgtt 781 gttgttaatg ggcattcctt cttctggtca gaaacctgtc cactgggcac agaacttatg 841 ttgttctcta tggagaacta aaagtatgag cgttaggaca ctattttaat tatttttaat 901 ttattaatat ttaaatatgt gaagctgagt taatttatgt aagtcatatt tatattttta 961 agaagtacca cttgaaacat tttatgtatt agttttgaaa taataatgga aagtggctat 1021 gcagtttgaa tatcctttgt ttcagagcca gatcatttct tggaaagtgt aggcttacct 1081 caaataaatg gctaacttat acatattttt aaagaaatat ttatattgta tttatataat 1141 gtataaatgg tttttatacc aataaatggc attttaaaaa attcagcaaa aaaaaaa Human IL-6 mRNA Transcript 2 (SEQ ID NO: 55) 1 gtctcaatat tagagtctca acccccaata aatataggac tggagatgtc tgaggctcat 61 tctgccctcg agcccaccgg gaacgaaaga gaagctctat ctcccctcca ggagcccagc 121 tatgaactcc ttctccacaa acatgtaaca agagtaacat gtgtgaaagc agcaaagagg 181 cactggcaga aaacaacctg aaccttccaa agatggctga aaaagatgga tgcttccaat 241 ctggattcaa tgaggagact tgcctggtga aaatcatcac tggtcttttg gagtttgagg 301 tatacctaga gtacctccag aacagatttg agagtagtga ggaacaagcc agagctgtgc 361 agatgagtac aaaagtcctg atccagttcc tgcagaaaaa ggcaaagaat ctagatgcaa 421 taaccacccc tgacccaacc acaaatgcca gcctgctgac gaagctgcag gcacagaacc 481 agtggctgca ggacatgaca actcatctca ttctgcgcag ctttaaggag ttcctgcagt 541 ccagcctgag ggctcttcgg caaatgtagc atgggcacct cagattgttg ttgttaatgg 601 gcattccttc ttctggtcag aaacctgtcc actgggcaca gaacttatgt tgttctctat 661 ggagaactaa aagtatgagc gttaggacac tattttaatt atttttaatt tattaatatt 721 taaatatgtg aagctgagtt aatttatgta agtcatattt atatttttaa gaagtaccac 781 ttgaaacatt ttatgtatta gttttgaaat aataatggaa agtggctatg cagtttgaat 841 atcctttgtt tcagagccag atcatttctt ggaaagtgta ggcttacctc aaataaatgg 901 ctaacttata catattttta aagaaatatt tatattgtat ttatataatg tataaatggt 961 ttttatacca ataaatggca ttttaaaaaa ttcagcaaaa aaaaaa Inhibitory Nucleic Acids An antisense nucleic acid molecule can be complementary to all or part of a non-coding region of the coding strand of a nucleotide sequence encoding an IL6R, sIL6gp130, or IL-6 protein. Non-coding regions (5' and 3' untranslated regions) are the 5' and 3' sequences that flank the coding region in a gene and are not translated into amino acids.
Based upon the sequences disclosed herein, one of skill in the art can easily choose and synthesize any of a number of appropriate antisense nucleic acids to target a nucleic acid encoding an IL6R, sIL6gp130, or IL-6 protein described herein. Antisense nucleic acids targeting a nucleic acid encoding an IL6R, sIL6gp130, or IL-6 protein can be designed using the software available at the Integrated DNA Technologies website.
An antisense nucleic acid can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides or more in length. An antisense oligonucleotide can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used.
Examples of modified nucleotides which can be used to generate an antisense nucleic acid include 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethy1-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethy1-2-thiouracil, beta-D-mannosylqueosine, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methy1-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methy1-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an anti sense orientation to a target nucleic acid of interest).
The antisense nucleic acid molecules described herein can be prepared in vitro and administered to a mammal, e.g., a human. Alternatively, they can be generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding an IL6R, sIL6gp130, or IL-6 protein to thereby inhibit expression, e.g., by inhibiting transcription and/or translation. The hybridization can be by conventional nucleotide complementarities to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA duplexes, through specific interactions in the major groove of the double helix.
The antisense nucleic acid molecules can be delivered to a mammalian cell using a vector (e.g., a lentivirus, a retrovirus, or an adenovirus vector).
An antisense nucleic acid can be an a-anomeric nucleic acid molecule. An a-anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual, 13-units, the strands run parallel to each other (Gaultier et al., Nucleic Acids Res. 15:6625-6641, 1987). The antisense nucleic acid can also comprise a 2'-0-methylribonucleotide (Inoue et al., Nucleic Acids Res. 15:6131-6148, 1987) or a chimeric RNA-DNA analog (Inoue et al., FEBS Lett. 215:327-330, 1987).

Exemplary antisense nucleic acids that are IL-6 receptor inhibitors are described in Keller et al., I Immunol. 154(8):4091-4098, 1995; and Jiang et al., Anticancer Res. 31(9):
2899-2906, 2011.
Another example of an inhibitory nucleic acid is a ribozyme that has specificity for a nucleic acid encoding an IL6R, sIL6gp130, or IL-6 protein (e.g., specificity for an IL6R, sIL6gp130, or IL-6 mRNA, e.g., specificity for any one of SEQ ID NOs: 50-55).
Ribozymes are catalytic RNA molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes (described in Haselhoff and Gerlach, Nature 334:585-591, 1988)) can be used to catalytically cleave mRNA transcripts to thereby inhibit translation of the protein encoded by the mRNA. A ribozyme having specificity for an IL6R, sIL6gp130, or IL-6 mRNA can be designed based upon the nucleotide sequence of any of the IL6R, sIL6gp130, or IL-6 mRNA sequences disclosed herein. For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in an IL6R, sIL6gp130, or IL-6 mRNA (see, e.g., U.S. Patent. Nos. 4,987,071 and 5,116,742).
Alternatively, a SMAD7 mRNA can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel et al., Science 261:1411-1418, 1993.
An inhibitory nucleic acid can also be a nucleic acid molecule that forms triple helical .. structures. For example, expression of an IL6R, sIL6gp130, or IL-6 polypeptide can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the gene encoding the IL6R, sIL6gp130, or IL-6 polypeptide (e.g., the promoter and/or enhancer, e.g., a sequence that is at least 1 kb, 2 kb, 3 kb, 4 kb, or 5 kb upstream of the transcription initiation start state) to form triple helical structures that prevent transcription of the gene in target cells. See generally Helene, Anticancer Drug Des. 6(6):569-84, 1991;
Helene, Ann.
N.Y. Acad. Sci. 660:27-36, 1992; and Maher, Bioassays 14(12):807-15, 1992.
In various embodiments, inhibitory nucleic acids can be modified at the base moiety, sugar moiety, or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acids can be modified to generate peptide nucleic acids (see, e.g., Hyrup et al., Bioorganic Medicinal Chem. 4(1):5-23, 1996). Peptide nucleic acids (PNAs) are nucleic acid mimics, e.g., DNA mimics, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs allows for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols (see, e.g., Perry-O'Keefe et al., Proc.
Natl. Acad. Sci.
U.S.A. 93:14670-675, 1996). PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication.
PNAs can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA
chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras can be generated which may combine the advantageous properties of PNA
and DNA. Such chimeras allow DNA recognition enzymes, e.g., RNAse H and DNA
polymerases, to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation.
The synthesis of PNA-DNA chimeras can be performed as described in Finn et al., Nucleic Acids Res. 24:3357-63, 1996. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry and modified nucleoside analogs. Compounds such as 5'-(4-methoxytrityl)amino-5'-deoxy-thymidine phosphoramidite can be used as a link between the PNA and the 5' end of DNA
(Mag et al., Nucleic Acids Res. 17:5973-88, 1989). PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5' PNA segment and a 3' DNA
segment (Finn et al., Nucleic Acids Res. 24:3357-63, 1996). Alternatively, chimeric molecules can be synthesized with a 5' DNA segment and a 3' PNA segment (Peterser et al., Bioorganic Med.
Chem. Lett. 5:1119-11124, 1975).
In some embodiments, the inhibitory nucleic acids can include other appended groups such as peptides, or agents facilitating transport across the cell membrane (see, Letsinger et al., Proc. Natl. Acad. Sci. U.S.A. 86:6553-6556, 1989; Lemaitre et al., Proc.
Natl. Acad. Sci.
U.S.A. 84:648-652, 1989; and WO 88/09810). In addition, the inhibitory nucleic acids can be modified with hybridization-triggered cleavage agents (see, e.g., Krol et al., Bio/Techniques 6:958-976, 1988) or intercalating agents (see, e.g., Zon, Pharm. Res. 5:539-549, 1988). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent, etc.
Another means by which expression of an IL6R, sIL6gp130, or IL-6 mRNA can be decreased in a mammalian cell is by RNA interference (RNAi). RNAi is a process in which mRNA is degraded in host cells. To inhibit an mRNA, double-stranded RNA
(dsRNA) corresponding to a portion of the gene to be silenced (e.g., a gene encoding an IL6R, sIL6gp130, or IL-6 polypeptide) is introduced into a mammalian cell. The dsRNA
is digested into 21-23 nucleotide-long duplexes called short interfering RNAs (or siRNAs), which bind to a nuclease complex to form what is known as the RNA-induced silencing complex (or RISC). The RISC targets the homologous transcript by base pairing interactions between one of the siRNA strands and the endogenous mRNA. It then cleaves the mRNA about nucleotides from the 3' terminus of the siRNA (see Sharp et al., Genes Dev.
15:485-490, 2001, and Hammond et al., Nature Rev. Gen. 2:110-119, 2001).
RNA-mediated gene silencing can be induced in a mammalian cell in many ways, e.g., by enforcing endogenous expression of RNA hairpins (see, Paddison et al., Proc. Natl.
Acad. Sci. U.S.A. 99:1443-1448, 2002) or, as noted above, by transfection of small (21-23 nt) dsRNA (reviewed in Caplen, Trends Biotech. 20:49-51, 2002). Methods for modulating gene expression with RNAi are described, e.g., in U.S. Patent No. 6,506,559 and US
2003/0056235, which are hereby incorporated by reference.
Standard molecular biology techniques can be used to generate siRNAs. Short interfering RNAs can be chemically synthesized, recombinantly produced, e.g., by expressing RNA from a template DNA, such as a plasmid, or obtained from commercial vendors, such as Dharmacon. The RNA used to mediate RNAi can include synthetic or modified nucleotides, such as phosphorothioate nucleotides. Methods of transfecting cells with siRNA
or with plasmids engineered to make siRNA are routine in the art.
The siRNA molecules used to decrease expression of an IL6R, sIL6gp130, or IL-6 mRNA can vary in a number of ways. For example, they can include a 3' hydroxyl group and strands of 21, 22, or 23 consecutive nucleotides. They can be blunt ended or include an overhanging end at either the 3' end, the 5' end, or both ends. For example, at least one strand of the RNA molecule can have a 3' overhang from about 1 to about 6 nucleotides (e.g., 1-5, 1-3, 2-4, or 3-5 nucleotides (whether pyrimidine or purine nucleotides) in length. Where both strands include an overhang, the length of the overhangs may be the same or different for each strand.

To further enhance the stability of the RNA duplexes, the 3' overhangs can be stabilized against degradation (by, e.g., including purine nucleotides, such as adenosine or guanosine nucleotides or replacing pyrimidine nucleotides by modified analogues (e.g., substitution of uridine 2-nucleotide 3' overhangs by 2'-deoxythymidine is tolerated and does not affect the efficiency of RNAi). Any siRNA can be used in the methods of decreasing an IL6R, sIL6gp130, or IL-6 mRNA, provided it has sufficient homology to the target of interest (e.g., a sequence present in any one of SEQ ID NOs: 50-55, e.g., a target sequence encompassing the translation start site or the first exon of the mRNA). There is no upper limit on the length of the siRNA that can be used (e.g., the siRNA can range from about 21 base pairs of the gene to the full length of the gene or more (e.g., about 20 to about 30 base pairs, about 50 to about 60 base pairs, about 60 to about 70 base pairs, about 70 to about 80 base pairs, about 80 to about 90 base pairs, or about 90 to about 100 base pairs).
Non-limiting examples of short interfering RNA (siRNA) that are IL-6 receptor inhibitors are described in Yi et al., Int. I Oncol. 41(1):310-316, 2012; and Shinriki et al., Cl/n. Can. Res. 15(17):5426-5434, 2009). Non-limiting examples of microRNAs that are IL-6 receptor inhibitors are described in miR34a (Li et al., Int. I Cl/n. Exp.
Pathol. 8(2):1364-1373, 2015) and miR-451 (Liu et al., Cancer Epidemiol. 38(1):85-92, 2014).
Non-limiting examples of aptamers that are IL-6 receptor inhibitors are described in Meyer et al., RNA Biol. 11(1):57-65, 2014; Meyer et al., RNA Biol. 9(1):67-80, 2012; and Mittelberger et al., RNA Biol. 12(9):1043-1053, 2015. Additional examples of inhibitory nucleic acids that are IL-6 receptor inhibitors are described in, e.g., WO
96/040157.
In certain embodiments, a therapeutically effective amount of an inhibitory nucleic acid targeting a nucleic acid encoding an IL6R, sIL6gp130, or IL-6 protein can be administered to a subject (e.g., a human subject) in need thereof.
In some embodiments, the inhibitory nucleic acid can be about 10 nucleotides to about 40 nucleotides (e.g., about 10 to about 30 nucleotides, about 10 to about 25 nucleotides, about 10 to about 20 nucleotides, about 10 to about 15 nucleotides, 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotides, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36 nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, or 40 nucleotides) in length. One skilled in the art will appreciate that inhibitory nucleic acids may comprise at least one modified nucleic acid at either the 5' or 3'end of DNA or RNA.
As is known in the art, the term "thermal melting point (Tm)" refers to the temperature, under defined ionic strength, pH, and inhibitory nucleic acid concentration, at which 50% of the inhibitory nucleic acids complementary to the target sequence hybridize to the target sequence at equilibrium. In some embodiments, an inhibitory nucleic acid can bind specifically to a target nucleic acid under stingent conditions, e.g., those in which the salt concentration is at least about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30 C. for short oligonucleotides (e.g., 10 to 50 nucleotide). Stringent conditions can also be achieved with the addition of destabilizing agents such as formamide.
In some embodiments of any of the inhibitory nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding any one of IL6R, sIL6gp130, or IL-6) with a Tm of greater than 20 C, greater than 22 C, greater than 24 C, greater than 26 C, greater than 28 C, greater than 30 C, greater than 32 C, greater than 34 C, greater than 36 C, greater than 38 C, greater than 40 C, greater than 42 C, greater than 44 C, greater than 46 C, greater than 48 C, greater than 50 C, greater than 52 C, greater than 54 C, greater than 56 C, greater than 58 C, greater than 60 C, greater than 62 C, greater than 64 C, greater than 66 C, greater than 68 C, greater than 70 C, greater than 72 C, greater than 74 C, greater than 76 C, greater than 78 C, or greater than 80 C, e.g., as measured in phosphate buffered saline using a UV spectrophotometer.
In some embodiments of any of the inhibitor nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding any one of IL6R, sIL6gp130, or IL-6) with a Tm of about 20 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, about 30 C, about 28 C, about 26 C, about 24 C, or about (inclusive); about 22 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, about 30 C, about 28 C, about 26 C, or about 24 C (inclusive); about 24 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, about 30 C, about 28 C, or about 26 C
(inclusive); about 26 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, about 30 C, or about (inclusive); about 28 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, or about 30 C
(inclusive); about 30 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, or about 32 C (inclusive);
about 32 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, or about 34 C (inclusive); about 34 C
to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, or about 36 C (inclusive); about 36 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, or about 38 C (inclusive);
about 38 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, .. about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, or about 40 C

(inclusive); about 40 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, or about 42 C (inclusive); about 42 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, or about 44 C (inclusive); about 44 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, or about 46 C (inclusive); about 46 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, or about 48 C (inclusive);
about 48 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, or about 50 C (inclusive); about 50 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, or about (inclusive); about 52 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, or about 54 C (inclusive); about 54 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, or about 56 C (inclusive); about 56 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, or about 58 C (inclusive); about 58 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, or about 60 C (inclusive); about 60 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, or about 62 C (inclusive); about 62 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, or about 64 C
(inclusive); about 64 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, or about 66 C (inclusive); about 66 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, or about 68 C (inclusive); about 68 C
to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, or about 70 C
(inclusive); about 70 C to about 80 C, about 78 C, about 76 C, about 74 C, or about 72 C
(inclusive); about 72 C to about 80 C, about 78 C, about 76 C, or about 74 C (inclusive);
about 74 C to about 80 C, about 78 C, or about 76 C (inclusive); about 76 C to about 80 C or about 78 C (inclusive); or about 78 C to about 80 C (inclusive), In some embodiments, the inhibitory nucleic acid can be formulated in a nanoparticle (e.g., a nanoparticle including one or more synthetic polymers, e.g., Patil et al., Pharmaceutical Nanotechnol. 367:195-203, 2009; Yang et al., ACS Appl. Mater.
Interfaces, doi: 10.1021/acsami.6b16556, 2017; Perepelyuk et al., Mol. Ther. Nucleic Acids 6:259-268, 2017). In some embodiments, the nanoparticle can be a mucoadhesive particle (e.g., nanoparticles having a positively-charged exterior surface) (Andersen et al., Methods Mot.
Biol. 555:77-86, 2009). In some embodiments, the nanoparticle can have a neutrally-charged exterior surface.
In some embodiments, the inhibitory nucleic acid can be formulated, e.g., as a liposome (Buyens et al., I Control Release 158(3): 362-370, 2012; Scarabel et al., Expert Op/n. Drug Deliv. 17:1-14, 2017), a micelle (e.g., a mixed micelle) (Tangsangasaksri et al., BioMacromolecules 17:246-255, 2016; Wu et al., Nanotechnology, doi:
10.1088/1361-6528/aa6519, 2017), a microemulsion (WO 11/004395), a nanoemulsion, or a solid lipid nanoparticle (Sahay et al., Nature Biotechnol. 31:653-658, 2013; and Lin et al., Nanomedicine 9(1):105-120, 2014). Additional exemplary structural features of inhibitory nucleic acids and formulations of inhibitory nucleic acids are described in US
2016/0090598.
In some embodiments, a pharmaceutical composition can include a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In some examples, a pharmaceutical composition consists of a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In certain embodiments, the sterile saline is a pharmaceutical grade saline. In certain embodiments, a pharmaceutical composition can include one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition includes one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and phosphate-buffered saline (PBS). In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) and sterile phosphate-buffered saline (PBS). In some examples, the sterile saline is a pharmaceutical grade PBS.
In certain embodiments, one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) may be admixed with pharmaceutically acceptable active and/or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions depend on a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.
Pharmaceutical compositions including one or more inhibitory nucleic acids encompass any pharmaceutically acceptable salts, esters, or salts of such esters. Non-limiting examples of pharmaceutical compositions include pharmaceutically acceptable salts of inhibitory nucleic acids. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.
Also provided herein are prodrugs that can include additional nucleosides at one or both ends of an inhibitory nucleic acid which are cleaved by endogenous nucleases within the body, to form the active inhibitory nucleic acid.
Lipid moieties can be used to formulate an inhibitory nucleic acid. In certain such methods, the inhibitory nucleic acid is introduced into preformed liposomes or lipoplexes made of mixtures of cationic lipids and neutral lipids. In certain methods, inhibitory nucleic acid complexes with mono- or poly-cationic lipids are formed without the presence of a neutral lipid. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to a particular cell or tissue in a mammal. In some examples, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to fat tissue in a mammal. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to muscle tissue.
In certain embodiments, pharmaceutical compositions provided herein comprise one or more inhibitory nucleic acid and one or more excipients. In certain such embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethyl cellulose and polyvinylpyrrolidone.

In some examples, a pharmaceutical composition provided herein includes liposomes and emulsions. Liposomes and emulsions can be used to formulate hydrophobic compounds.
In some examples, certain organic solvents such as dimethylsulfoxide are used.
In some examples, a pharmaceutical composition provided herein includes one or more tissue-specific delivery molecules designed to deliver one or more inhibitory nucleic acids to specific tissues or cell types in a mammal. For example, a pharmaceutical composition can include liposomes coated with a tissue-specific antibody.
In some embodiments, a pharmaceutical composition provided herein can include a co-solvent system. Examples of such co-solvent systems include benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. A non-limiting example of such a co-solvent system is the VPD co-solvent system, which is a solution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80 and 65% w/v polyethylene glycol 300. As can be appreciated, other surfactants may be used instead of Polysorbate 80TM; the fraction size of polyethylene glycol may be varied;
other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone;
and other sugars or polysaccharides may substitute for dextrose.
In some examples, a pharmaceutical composition can be formulated for oral administration. In some examples, pharmaceutical compositions are formulated for buccal administration.
In some examples, a pharmaceutical composition is formulated for administration by injection (e.g., intravenous, subcutaneous, intramuscular, etc.). In some of these embodiments, a pharmaceutical composition includes a carrier and is formulated in aqueous solution, such as water or physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. In some examples, other ingredients are included (e.g., ingredients that aid in solubility or serve as preservatives).
In some examples, injectable suspensions are prepared using appropriate liquid carriers, suspending agents, and the like. Some pharmaceutical compositions for injection are formulated in unit dosage form, e.g., in ampoules or in multi-dose containers. Some pharmaceutical compositions for injection are suspensions, solutions, or emulsions in oily or aqueous vehicles, and may .. contain formulatory agents such as suspending, stabilizing, and/or dispersing agents.
Solvents suitable for use in pharmaceutical compositions for injection include, but are not limited to, lipophilic solvents and fatty oils, such as sesame oil, synthetic fatty acid esters, such as ethyl oleate or triglycerides, and liposomes.

Antibodies In some embodiments, the IL-6 receptor inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to IL-6. In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to IL-6 receptor (e.g., one or both of IL6R and sIL6gp130).
In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc, a VHEI domain, a VNAR domain, a (scFv)2, a minibody, or a BiTE. In some embodiments, an antibody can be a DVD-Ig, and a dual-affinity re-targeting antibody (DART), a triomab, kih IgG with a common LC, a crossmab, an ortho-Fab IgG, a 2-in-1-IgG, IgG-ScFv, seFv2-Fe, a bi-nanobody, tanden antibody, a DART-Fe, a scFv-HAS-scFv, DNL-Fab3, DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, la-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG (L,H)-Fc, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH
IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody, nanobody-HSA, a diabody, a TandAb, scDiabody, scDiabody-CH3, Diabody-CH3, Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab')2-seFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody, dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HAS, tandem scFv, IgG-IgG, Cov-X-Body, and seFvl-PEG-scFv2.
Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab')2 fragment, and a Fab' fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgGl, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgGl, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgAl or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgAl or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).

In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Patent Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895;
US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US
2017/0002082, each of which is incorporated by reference in its entirety.
In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of tocilizumab (artlizumab, Actemrag; Sebba, Am. I Health Syst.
Pharm. 65(15):1413-1418, 2008; Tanaka et al., FEBS Letters 585(23):3699-3709, 2011;
Nishimoto et al., Arthritis Rheum. 50:1761-1769, 2004; Yokota et al., Lancet 371(9617):998-1006, 2008; Emery et al., Ann. Rheum. Dis. 67(11):1516-1523, 2008; Roll et al., Arthritis Rheum. 63(5):1255-1264, 2011); lazakizumab (BM5945429; ALD518, a humanized monoclonal antibody that binds circulating IL-6 cytokine rather than the IL-6 receptor, blocking both classic signaling and trans-signaling (Weinblatt, Michael E., et al. "The Efficacy and Safety of Subcutaneous Clazakizumab in Patients With Moderate-to-Severe Rheumatoid Arthritis and an Inadequate Response to Methotrexate: Results From a Multinational, Phase IIb, Randomized, Double-Blind, Placebo/Active-Controlled, Dose-Ranging Study." Arthritis & Rheumatology 67.10 (2015): 2591-2600.)); sarilumab (REGN88 or SAR153191; Huizinga et al., Ann. Rheum. Dis. 73(9):1626-1634, 2014; Sieper et al., Ann.
Rheum. Dis.74(6):1051-1057, 2014; Cooper, Immunotherapy 8(3): 249-250, 2016);

(Hartman et al., PLosOne 11(12):e0167195, 2016; Fujita et al., Biochim.
Biophys. Acta.
10:3170-80, 2014; Okazaki et al., Immunol. Lett. 84(3):231-40, 2002; Noguchi-Sasaki et al., BMC Cancer 16:270, 2016; Ueda et al., Sci. Rep. 3:1196, 2013); rhPM-1 (MRA;
Nishimoto et al., Blood 95: 56-61, 2000; Nishimoto et al., Blood 106: 2627-2632, 2005;
Nakahara et al., Arthritis Rheum. 48(6): 1521-1529, 2003); NI-1201 (Lacroix et al., I Biol.
Chem.
290(45):26943-26953, 2015); EBI-029 (Schmidt et al., Eleven Biotherapeutics Poster #B0200, 2014). In some embodiments, the antibody is a nanobody (e.g., ALX-0061 (Van Roy et al., Arthritis Res. Ther. 17: 135, 2015; Kim et al., Arch. Pharm. Res.
38(5):575-584, 2015)). In some embodiments, the antibody is NRI or a variant thereof (Adachi et al., Mol.
Ther. 11(1):5262-263, 2005; Hoshino et al., Can. Res. 67(3): 871-875, 2007).
In some embodiments, the antibody is PF-04236921 (Pfizer) (Wallace et al., Ann. Rheum.
Dis.
76(3):534-542, 2017).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (KD) of less than 1 x 10-5M (e.g., less than 0.5 x 10-5M, less than 1 x 10' M, less than 0.5 x 10' M, less than 1 x 107M, less than 0.5 x 10-7M, less than 1 x 10-8M, less than 0.5 x 10-8M, less than 1 x 10-9M, less than 0.5 x 10-9M, less than 1 x 10' M, less than 0.5 x 10-1 M, less than 1 x 10-11M, less than 0.5 x 10-11M, or less than 1 x 1012M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a KD of about lx 10-12M to about lx 10-5M, about 0.5 x 10-5M, about lx 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8 M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-1 M, about 0.5 x 10-1 M, about 1 x 10-11M, or about 0.5 x 10-11M (inclusive); about 0.5 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-1 M, about 0.5 x 10' M, or about 1 x 10-11M (inclusive); about 1 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-1 M, or about 0.5 x 10-1 M (inclusive); about 0.5 x 10-10 M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, or about 1 x 10' M
(inclusive);
about 1 x 10' M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9 M, or about 0.5 x 10-9M (inclusive); about 0.5 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 107M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, or about 1 x 10-9M (inclusive); about 1 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, about 1 x 10-8M, or about 0.5 x 10-8M (inclusive); about 0.5 x 10-8M to about 1 x 10-5 M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 107M, about 0.5 x 10-7 M, or about 1 x 10-8M (inclusive); about 1 x 10-8M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10-6M, about 1 x 10-7M, or about 0.5 x 10-7M
(inclusive);
about 0.5 x 10-7M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, or about 1 x 107M (inclusive); about 1 x 10-7 M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, or about 0.5 x 10' M (inclusive); about 0.5 x 10' M to about 1 x 10-5M, about 0.5 x 10-5 M, or about 1 x 10' M (inclusive); about 1 x 10' M to about 1 x 10-5M or about 0.5 x 10-5 M (inclusive); or about 0.5 x 10-5M to about 1 x 10-5M
(inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Koff-of about lx 10' s-1 to about lx 10-3 s-1, about 0.5 x 10-3 s-1, about lx 10-4 s"
1, about 0.5 x 10-4 s-1, about 1 x 10-5 s-1, or about 0.5 x 10-5 s-1 (inclusive); about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, about 0.5 x 10-4 s-1, or about 1 x 10-5 s-1 (inclusive); about 1 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s"
',or about 0.5 x 10-4 s-1 (inclusive); about 0.5 x 10-4 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 or about 1 x 10-4 s-1 (inclusive); about 1 x 10-4 s-lto about 1 x 10-3 s-1, or about 0.5 x 10-3 s-1 (inclusive); or about 0.5 x 10-5 s-lto about 1 x 10-3 s-1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Km, of about lx 102 M's' to about lx 106 M's', about 0.5 x 106 M's', about 1 x 105M-1s-1, about 0.5 x 105 M's', about 1 x 104 m-1S-1, about 0.5 x 104 M's', about 1 x 103 M's', or about 0.5 x 103 M-1s-1 (inclusive); about 0.5 x 103 M-1s-1 to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105M-1s-1, about 0.5 x 105 M's', about 1 x 104 M's', about 0.5 x 104 M's', or about 1 x 103 M-1s-1 (inclusive); about 1 x 103 M-1s-lto about 1 x 106 M-1s-1, about 0.5 x 106 M's', about 1 x 105M-1s-1, about 0.5 x 105 M's', about 1 x 104 M's', or about 0.5 x 104 m-ls-1 (inclusive); about 0.5 x 104 m-is-lto about 1 x 106M-1s-1, about 0.5 x 106 M's', about 1 x i05 M's', about 0.5 x 105 M's', or about 1 x 104 M-1s-1 (inclusive);
about 1 x 104 M's' to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105M-is-i, or about 0.5 x 105 M's' (inclusive); about 0.5 x 105 M-1s-lto about 1 x 106M-1s-1, about 0.5 x 106 M's', or about 1 x 105 M's' (inclusive); about 1 x 105M-1s-1 to about 1 x 106 M's', or about 0.5 x 106 m-ls-1 (inclusive); or about 0.5 x 106 to about 1 X 106 M's' (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
Fusion Proteins In some embodiments, the IL-6 receptor inhibitor is a fusion protein, a soluble receptor, or a peptide (see e.g., U.S. Patent No. 5,591,827). In some embodiments, the IL-6 receptor fusion protein comprises or consists of soluble gp130 (Jostock et al., Eur.

Biochem. 268(1):160-167, 2001; Richards etal., Arthritis Rheum. 54(5):1662-1672, 2006;
Rose-John etal., Exp. Op/n. Ther. Targets 11(5):613-624, 2007).
In some embodiments, the IL-6 receptor fusion protein comprises or consists of FE999301 (Jostock etal., Eur. I Biochem. 268(1):160-167, 2001) or sgp130Fc protein (Jones et al., I Cl/n. Invest. 121(9):3375-3383, 2011). In some embodiments, the IL-6 receptor inhibitor is a peptide (e.g., S7 (Su etal., Cancer Res. 65(11):4827-4835, 2005). In some embodiments, the IL-6 receptor inhibitor is a triterpenoid saponin (e.g., chikusetsuaponin IVa butyl ester (CS-Iva-Be) (Yang et al., Mol. Cancer. Ther. 15(6):1190-200, 2016).
Small Molecules In some embodiments, the IL-6 receptor inhibitor is a small molecule (see, e.g., U.S.
Patent No. 9,409,990). In some embodiments, the small molecule is LMT-28 (Hong et al., Immunol. 195(1): 237-245, 2015); ERBA (Enomoto et al .,Biochem. Biophys. Res.
Commun.
323:1096-1102, 2004; Boos etal., I Nat. Prod. 75(4):661-668, 2012), ERBF (TB-2-081) (Hayashi etal., I Pharmacol. Exp. Ther. 303:104-109, 2002; Vardanyan etal., Pain 151(2):257-265, 2010; Kino etal., I Allergy Cl/n. Immunol. 120(2):437-444, 2007), or a variant thereof.
Immune Modulatory Agents As used herein, the term "immune modulatory agentomodifier " refers to an agent that is a CD40/CD40 inhibitor (as defined herein), a CD3 inhibitor (as defined herein), a CD14 inhibitor (as defined agent), a CD20 inhibitor (as defined herein), a CD25 inhibitor (as defined herein), a CD28 inhibitor (as defined herein), a CD49 inhibitor (as defined herein), or a CD89 inhibitor. Examples of immune modulatory agents are described herein.
Additional examples of immune modulatory agents are known in the art.
CD40/CD4OL Inhibitors The term "CD40/CD4OL inhibitors" refers to an agent which decreases CD40 or CD4OL (CD154) expression and/or the ability of CD40 to bind to CD4OL (CD154).
CD40 is a costimulatory receptor that binds to its ligand, CD4OL (CD154).
In some embodiments, the CD40/CD4OL inhibitor can decrease the binding between CD40 and CD4OL by blocking the ability of CD40 to interact with CD4OL. In some embodiments, the CD40/CD4OL inhibitor can decrease the binding between CD40 and CD4OL by blocking the ability of CD4OL to interact with CD40. In some embodiments, the CD40/CD4OL inhibitor decreases the expression of CD40 or CD4OL. In some embodiments, the CD40/CD4OL inhibitor decreases the expression of CD40. In some embodiments, the CD40/CD4OL inhibitor decreases the expression of CD4OL.
In some embodiments, the CD40/CD4OL inhibitor is an inhibitory nucleic acid, an antibody or an antigen-binding fragment thereof, a fusion protein, or a small molecule. In some embodiments, the inhibitory nucleic acid is a small interfering RNA, an antisense nucleic acid, an aptamer, or a microRNA. Exemplary CD40/CD4OL inhibitors are described herein. Additional examples of CD40/CD4OL inhibitors are known in the art.
Exemplary aspects of different inhibitory nucleic acids are described below.
Any of the examples of inhibitory nucleic acids that can decrease expression of CD40 or CD4OL
mRNA in a mammalian cell can be synthesized in vitro. Inhibitory nucleic acids that can decrease the expression of CD40 or CD4OL mRNA in a mammalian cell include antisense nucleic acid molecules, i.e., nucleic acid molecules whose nucleotide sequence is complementary to all or part of a CD40 or CD4OL mRNA (e.g., complementary to all or a part of any one of SEQ ID NOs: 56-61).
Human CD40 mRNA (Variant 1) NM 001250.5 (SEQ ID NO: 56) 1 tttcctgggc ggggccaagg ctggggcagg ggagtcagca gaggcctcgc tcgggcgccc 61 agtggtcctg ccgcctggtc tcacctcgct atggttcgtc tgcctctgca gtgcgtcctc 121 tggggctgct tgctgaccgc tgtccatcca gaaccaccca ctgcatgcag agaaaaacag 181 tacctaataa acagtcagtg ctgttctttg tgccagccag gacagaaact ggtgagtgac 241 tgcacagagt tcactgaaac ggaatgcctt ccttgcggtg aaagcgaatt cctagacacc 301 tggaacagag agacacactg ccaccagcac aaatactgcg accccaacct agggcttcgg 361 gtccagcaga agggcacctc agaaacagac accatctgca cctgtgaaga aggctggcac 421 tgtacgagtg aggcctgtga gagctgtgtc ctgcaccgct catgctcgcc cggctttggg 481 gtcaagcaga ttgctacagg ggtttctgat accatctgcg agccctgccc agtcggcttc 541 ttctccaatg tgtcatctgc tttcgaaaaa tgtcaccctt ggacaagctg tgagaccaaa 601 gacctggttg tgcaacaggc aggcacaaac aagactgatg ttgtctgtgg tccccaggat 661 cggctgagag ccctggtggt gatccccatc atcttcggga tcctgtttgc catcctcttg 721 gtgctggtct ttatcaaaaa ggtggccaag aagccaacca ataaggcccc ccaccccaag 781 caggaacccc aggagatcaa ttttcccgac gatcttcctg gctccaacac tgctgctcca 841 gtgcaggaga ctttacatgg atgccaaccg gtcacccagg aggatggcaa agagagtcgc 901 atctcagtgc aggagagaca gtgaggctgc acccacccag gagtgtggcc acgtgggcaa 961 acaggcagtt ggccagagag cctggtgctg ctgctgctgt ggcgtgaggg tgaggggctg 1021 gcactgactg ggcatagctc cccgcttctg cctgcacccc tgcagtttga gacaggagac 1081 ctggcactgg atgcagaaac agttcacctt gaagaacctc tcacttcacc ctggagccca 1141 tccagtctcc caacttgtat taaagacaga ggcagaagtt tggtggtggt ggtgttgggg 1201 tatggtttag taatatccac cagaccttcc gatccagcag tttggtgccc agagaggcat 1261 catggtggct tccctgcgcc caggaagcca tatacacaga tgcccattgc agcattgttt 1321 gtgatagtga acaactggaa gctgcttaac tgtccatcag caggagactg gctaaataaa 1381 attagaatat atttatacaa cagaatctca aaaacactgt tgagtaagga aaaaaaggca 1441 tgctgctgaa tgatgggtat ggaacttttt aaaaaagtac atgcttttat gtatgtatat 1501 tgcctatgga tatatgtata aatacaatat gcatcatata ttgatataac aagggttctg 1561 gaagggtaca cagaaaaccc acagctcgaa gagtggtgac gtctggggtg gggaagaagg 1621 gtctggggg Human CD40 mRNA (Variant 2) NM 152854.3 (SEQ ID NO: 57) 1 tttcctgggc ggggccaagg ctggggcagg ggagtcagca gaggcctcgc tcgggcgccc 61 agtggtcctg ccgcctggtc tcacctcgct atggttcgtc tgcctctgca gtgcgtcctc 121 tggggctgct tgctgaccgc tgtccatcca gaaccaccca ctgcatgcag agaaaaacag 181 tacctaataa acagtcagtg ctgttctttg tgccagccag gacagaaact ggtgagtgac 241 tgcacagagt tcactgaaac ggaatgcctt ccttgcggtg aaagcgaatt cctagacacc 301 tggaacagag agacacactg ccaccagcac aaatactgcg accccaacct agggcttcgg 361 gtccagcaga agggcacctc agaaacagac accatctgca cctgtgaaga aggctggcac 421 tgtacgagtg aggcctgtga gagctgtgtc ctgcaccgct catgctcgcc cggctttggg 481 gtcaagcaga ttgctacagg ggtttctgat accatctgcg agccctgccc agtcggcttc 541 ttctccaatg tgtcatctgc tttcgaaaaa tgtcaccctt ggacaaggtc cccaggatcg 601 gctgagagcc ctggtggtga tccccatcat cttcgggatc ctgtttgcca tcctcttggt 661 gctggtcttt atcaaaaagg tggccaagaa gccaaccaat aaggcccccc accccaagca 721 ggaaccccag gagatcaatt ttcccgacga tcttcctggc tccaacactg ctgctccagt 781 gcaggagact ttacatggat gccaaccggt cacccaggag gatggcaaag agagtcgcat 841 ctcagtgcag gagagacagt gaggctgcac ccacccagga gtgtggccac gtgggcaaac 901 aggcagttgg ccagagagcc tggtgctgct gctgctgtgg cgtgagggtg aggggctggc 961 actgactggg catagctccc cgcttctgcc tgcacccctg cagtttgaga caggagacct 1021 ggcactggat gcagaaacag ttcaccttga agaacctctc acttcaccct ggagcccatc 1081 cagtctccca acttgtatta aagacagagg cagaagtttg gtggtggtgg tgttggggta 1141 tggtttagta atatccacca gaccttccga tccagcagtt tggtgcccag agaggcatca 1201 tggtggcttc cctgcgccca ggaagccata tacacagatg cccattgcag cattgtttgt 1261 gatagtgaac aactggaagc tgcttaactg tccatcagca ggagactggc taaataaaat 1321 tagaatatat ttatacaaca gaatctcaaa aacactgttg agtaaggaaa aaaaggcatg 1381 ctgctgaatg atgggtatgg aactttttaa aaaagtacat gcttttatgt atgtatattg 1441 cctatggata tatgtataaa tacaatatgc atcatatatt gatataacaa gggttctgga 1501 agggtacaca gaaaacccac agctcgaaga gtggtgacgt ctggggtggg gaagaagggt 1561 ctggggg Human CD40 mRNA (Variant 3) NM 001302753.1 (SEQ ID NO: 58) 1 tttcctgggc ggggccaagg ctggggcagg ggagtcagca gaggcctcgc tcgggcgccc 61 agtggtcctg ccgcctggtc tcacctcgct atggttcgtc tgcctctgca gtgcgtcctc 121 tggggctgct tgctgaccgc tgtccatcca gaaccaccca ctgcatgcag agaaaaacag 181 tacctaataa acagtcagtg ctgttctttg tgccagccag gacagaaact ggtgagtgac 241 tgcacagagt tcactgaaac ggaatgcctt ccttgcggtg aaagcgaatt cctagacacc 301 tggaacagag agacacactg ccaccagcac aaatactgcg accccaacct agggcttcgg 361 gtccagcaga agggcacctc agaaacagac accatctgca cctgtgaaga aggctggcac 421 tgtacgagtg aggcctgtga gagctgtgtc ctgcaccgct catgctcgcc cggctttggg 481 gtcaagcaga ttgctacagg ggtttctgat accatctgcg agccctgccc agtcggcttc 541 ttctccaatg tgtcatctgc tttcgaaaaa tgtcaccctt ggacaagctg tgagaccaaa 601 gacctggttg tgcaacaggc aggcacaaac aagactgatg ttgtctgtgg tgagtcctgg 661 acaatgggcc ctggagaaag cctaggaagg tccccaggat cggctgagag ccctggtggt 721 gatccccatc atcttcggga tcctgtttgc catcctcttg gtgctggtct ttatcaaaaa 781 ggtggccaag aagccaacca ataaggcccc ccaccccaag caggaacccc aggagatcaa 841 ttttcccgac gatcttcctg gctccaacac tgctgctcca gtgcaggaga ctttacatgg 901 atgccaaccg gtcacccagg aggatggcaa agagagtcgc atctcagtgc aggagagaca 961 gtgaggctgc acccacccag gagtgtggcc acgtgggcaa acaggcagtt ggccagagag 1021 cctggtgctg ctgctgctgt ggcgtgaggg tgaggggctg gcactgactg ggcatagctc 1081 cccgcttctg cctgcacccc tgcagtttga gacaggagac ctggcactgg atgcagaaac 1141 agttcacctt gaagaacctc tcacttcacc ctggagccca tccagtctcc caacttgtat 1201 taaagacaga ggcagaagtt tggtggtggt ggtgttgggg tatggtttag taatatccac 1261 cagaccttcc gatccagcag tttggtgccc agagaggcat catggtggct tccctgcgcc 1321 caggaagcca tatacacaga tgcccattgc agcattgttt gtgatagtga acaactggaa 1381 gctgcttaac tgtccatcag caggagactg gctaaataaa attagaatat atttatacaa 1441 cagaatctca aaaacactgt tgagtaagga aaaaaaggca tgctgctgaa tgatgggtat 1501 ggaacttttt aaaaaagtac atgcttttat gtatgtatat tgcctatgga tatatgtata 1561 aatacaatat gcatcatata ttgatataac aagggttctg gaagggtaca cagaaaaccc 1621 acagctcgaa gagtggtgac gtctggggtg gggaagaagg gtctggggg Human CD40 mRNA (Variant 5) NM_001322421.1 (SEQ ID NO: 59) 1 tttcctgggc ggggccaagg ctggggcagg ggagtcagca gaggcctcgc tcgggcgccc 61 agtggtcctg ccgcctggtc tcacctcgct atggttcgtc tgcctctgca gtgcgtcctc 121 tggggctgct tgctgaccgc tgtccatcca gaaccaccca ctgcatgcag agaaaaacag 181 tacctaataa acagtcagtg ctgttctttg tgccagccag gacagaaact ggtgagtgac 241 tgcacagagt tcactgaaac ggaatgcctt ccttgcggtg aaagcgaatt cctagacacc 301 tggaacagag agacacactg ccaccagcac aaatactgcg accccaacct agggcttcgg 361 gtccagcaga agggcacctc agaaacagac accatctgca cctgtgaaga aggctggcac 421 tgtacgagtg aggcctgtga gagctgtgtc ctgcaccgct catgctcgcc cggctttggg 481 gtcaagcaga ttgctacagg ggtttctgat accatctgcg agccctgccc agtcggcttc 541 ttctccaatg tgtcatctgc tttcgaaaaa tgtcaccctt ggacaagctg tgagaccaaa 601 gacctggttg tgcaacaggc aggcacaaac aagactgatg ttgtctgtgg tccccaggat 661 cggctgagag ccctggtggt gatccccatc atcttcggga tcctgtttgc catcctcttg 721 gtgctggtct ttatcagtga gtcctcagaa aaggtggcca agaagccaac caataaggcc 781 ccccacccca agcaggaacc ccaggagatc aattttcccg acgatcttcc tggctccaac 841 actgctgctc cagtgcagga gactttacat ggatgccaac cggtcaccca ggaggatggc 901 aaagagagtc gcatctcagt gcaggagaga cagtgaggct gcacccaccc aggagtgtgg 961 ccacgtgggc aaacaggcag ttggccagag agcctggtgc tgctgctgct gtggcgtgag 1021 ggtgaggggc tggcactgac tgggcatagc tccccgcttc tgcctgcacc cctgcagttt 1081 gagacaggag acctggcact ggatgcagaa acagttcacc ttgaagaacc tctcacttca 1141 ccctggagcc catccagtct cccaacttgt attaaagaca gaggcagaag tttggtggtg 1201 gtggtgttgg ggtatggttt agtaatatcc accagacctt ccgatccagc agtttggtgc 1261 ccagagaggc atcatggtgg cttccctgcg cccaggaagc catatacaca gatgcccatt 1321 gcagcattgt ttgtgatagt gaacaactgg aagctgctta actgtccatc agcaggagac 1381 tggctaaata aaattagaat atatttatac aacagaatct caaaaacact gttgagtaag 1441 gaaaaaaagg catgctgctg aatgatgggt atggaacttt ttaaaaaagt acatgctttt 1501 atgtatgtat attgcctatg gatatatgta taaatacaat atgcatcata tattgatata 1561 acaagggttc tggaagggta cacagaaaac ccacagctcg aagagtggtg acgtctgggg 1621 tggggaagaa gggtctgggg g Human CD40 mRNA (Variant 6) NM_001322422.1 (SEQ ID NO: 60) 1 tttcctgggc ggggccaagg ctggggcagg ggagtcagca gaggcctcgc tcgggcgccc 61 agtggtcctg ccgcctggtc tcacctcgct atggttcgtc tgcctctgca gtgcgtcctc 121 tggggctgct tgctgaccgc tgtccatcca gaaccaccca ctgcatgcag agaaaaacag 181 tacctaataa acagtcagtg ctgttctttg tgccagccag gacagaaact ggtgagtgac 241 tgcacagagt tcactgaaac ggaatgcctt ccttgcggtg aaagcgaatt cctagacacc 301 tggaacagag agacacactg ccaccagcac aaatactgcg accccaacct agggcttcgg 361 gtccagcaga agggcacctc agaaacagac accatctgca cctgtgaaga aggctggcac 421 tgtacgagtg aggcctgtga gagctgtgtc ctgcaccgct catgctcgcc cggctttggg 481 gtcaagcaga ttggtcccca ggatcggctg agagccctgg tggtgatccc catcatcttc 541 gggatcctgt ttgccatcct cttggtgctg gtctttatca aaaaggtggc caagaagcca 601 accaataagg ccccccaccc caagcaggaa ccccaggaga tcaattttcc cgacgatctt 661 cctggctcca acactgctgc tccagtgcag gagactttac atggatgcca accggtcacc 721 caggaggatg gcaaagagag tcgcatctca gtgcaggaga gacagtgagg ctgcacccac 781 ccaggagtgt ggccacgtgg gcaaacaggc agttggccag agagcctggt gctgctgctg 841 ctgtggcgtg agggtgaggg gctggcactg actgggcata gctccccgct tctgcctgca 901 cccctgcagt ttgagacagg agacctggca ctggatgcag aaacagttca ccttgaagaa 961 cctctcactt caccctggag cccatccagt ctcccaactt gtattaaaga cagaggcaga 1021 agtttggtgg tggtggtgtt ggggtatggt ttagtaatat ccaccagacc ttccgatcca 1081 gcagtttggt gcccagagag gcatcatggt ggcttccctg cgcccaggaa gccatataca 1141 cagatgccca ttgcagcatt gtttgtgata gtgaacaact ggaagctgct taactgtcca 1201 tcagcaggag actggctaaa taaaattaga atatatttat acaacagaat ctcaaaaaca 1261 ctgttgagta aggaaaaaaa ggcatgctgc tgaatgatgg gtatggaact ttttaaaaaa 1321 gtacatgctt ttatgtatgt atattgccta tggatatatg tataaataca atatgcatca 1381 tatattgata taacaagggt tctggaaggg tacacagaaa acccacagct cgaagagtgg 1441 tgacgtctgg ggtggggaag aagggtctgg ggg Human CD154 (CD4OL) mRNA NM 000074.2 (SEQ ID NO: 61) 1 actttgacag tcttctcatg ctgcctctgc caccttctct gccagaagat accatttcaa 61 ctttaacaca gcatgatcga aacatacaac caaacttctc cccgatctgc ggccactgga 121 ctgcccatca gcatgaaaat ttttatgtat ttacttactg tttttcttat cacccagatg 181 attgggtcag cactttttgc tgtgtatctt catagaaggt tggacaagat agaagatgaa 241 aggaatcttc atgaagattt tgtattcatg aaaacgatac agagatgcaa cacaggagaa 301 agatccttat ccttactgaa ctgtgaggag attaaaagcc agtttgaagg ctttgtgaag 361 gatataatgt taaacaaaga ggagacgaag aaagaaaaca gctttgaaat gcaaaaaggt 421 gatcagaatc ctcaaattgc ggcacatgtc ataagtgagg ccagcagtaa aacaacatct 481 gtgttacagt gggctgaaaa aggatactac accatgagca acaacttggt aaccctggaa 541 aatgggaaac agctgaccgt taaaagacaa ggactctatt atatctatgc ccaagtcacc 601 ttctgttcca atcgggaagc ttcgagtcaa gctccattta tagccagcct ctgcctaaag 661 tcccccggta gattcgagag aatcttactc agagctgcaa atacccacag ttccgccaaa 721 ccttgcgggc aacaatccat tcacttggga ggagtatttg aattgcaacc aggtgcttcg 781 gtgtttgtca atgtgactga tccaagccaa gtgagccatg gcactggctt cacgtccttt 841 ggcttactca aactctgaac agtgtcacct tgcaggctgt ggtggagctg acgctgggag 901 tcttcataat acagcacagc ggttaagccc accccctgtt aactgcctat ttataaccct 961 aggatcctcc ttatggagaa ctatttatta tacactccaa ggcatgtaga actgtaataa 1021 gtgaattaca ggtcacatga aaccaaaacg ggccctgctc cataagagct tatatatctg 1081 aagcagcaac cccactgatg cagacatcca gagagtccta tgaaaagaca aggccattat 1141 gcacaggttg aattctgagt aaacagcaga taacttgcca agttcagttt tgtttctttg 1201 cgtgcagtgt ctttccatgg ataatgcatt tgatttatca gtgaagatgc agaagggaaa 1261 tggggagcct cagctcacat tcagttatgg ttgactctgg gttcctatgg ccttgttgga 1321 gggggccagg ctctagaacg tctaacacag tggagaaccg aaaccccccc cccccccccg 1381 ccaccctctc ggacagttat tcattctctt tcaatctctc tctctccatc tctctctttc 1441 agtctctctc tctcaacctc tttcttccaa tctctctttc tcaatctctc tgtttccctt 1501 tgtcagtctc ttccctcccc cagtctctct tctcaatccc cctttctaac acacacacac 1561 acacacacac acacacacac acacacacac acacacacac agagtcaggc cgttgctagt 1621 cagttctctt ctttccaccc tgtccctatc tctaccacta tagatgaggg tgaggagtag 1681 ggagtgcagc cctgagcctg cccactcctc attacgaaat gactgtattt aaaggaaatc 1741 tattgtatct acctgcagtc tccattgttt ccagagtgaa cttgtaatta tcttgttatt 1801 tattttttga ataataaaga cctcttaaca ttaa Inhibitory Nucleic Acids An antisense nucleic acid molecule can be complementary to all or part of a non-coding region of the coding strand of a nucleotide sequence encoding a CD40 or protein. Non-coding regions (5' and 3' untranslated regions) are the 5' and 3' sequences that flank the coding region in a gene and are not translated into amino acids.
Based upon the sequences disclosed herein, one of skill in the art can easily choose and synthesize any of a number of appropriate antisense nucleic acids to target a nucleic acid encoding a CD40 or CD4OL protein described herein. Antisense nucleic acids targeting a nucleic acid encoding a CD40 or CD4OL protein can be designed using the software available at the Integrated DNA Technologies website.

An antisense nucleic acid can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides or more in length. An antisense oligonucleotide can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid can be chemically synthesized using naturally-occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine-substituted nucleotides can be used.
Examples of modified nucleotides which can be used to generate an antisense nucleic acid include 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethy1-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethy1-2-thiouracil, beta-D-mannosylqueosine, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methy1-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methy1-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an anti sense orientation to a target nucleic acid of interest).
The antisense nucleic acid molecules described herein can be prepared in vitro and administered to a mammal, e.g., a human, using any of the devices described herein.
Alternatively, they can be generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding a CD40 or CD4OL protein to thereby inhibit expression, e.g., by inhibiting transcription and/or translation. The hybridization can be by conventional nucleotide complementarities to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA duplexes, through specific interactions in the major groove of the double helix. The antisense nucleic acid molecules can be delivered to a mammalian cell using a vector (e.g., a lentivirus, a retrovirus, or an adenovirus vector).

An anti sense nucleic acid can be an a-anomeric nucleic acid molecule. An a-anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual, 13-units, the strands run parallel to each other (Gaultier et al., Nucleic Acids Res. 15:6625-6641, 1987). The antisense nucleic acid can also comprise a 2'-0-methylribonucleotide (Inoue et al., Nucleic Acids Res. 15:6131-6148, 1987) or a chimeric RNA-DNA analog (Inoue et al., FEBS Lett. 215:327-330, 1987).
Some exemplary antisense nucleic acids that are CD40 or CD4OL inhibitors are described, e.g., in U.S. Patent Nos. 6,197,584 and 7,745,609; Gao et al., Gut 54(1):70-77, 2005; Arranz et al., I Control Release 165(3):163-172, 2012; Donner et al., Mol. Ther.
Nucleic Acids 4:e265, 2015.
Another example of an inhibitory nucleic acid is a ribozyme that has specificity for a nucleic acid encoding a CD40 or CD4OL protein (e.g., specificity for a CD40 or mRNA, e.g., specificity for any one of SEQ ID NOs: 56-61). Ribozymes are catalytic RNA
molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes (described in Haselhoff and Gerlach, Nature 334:585-591, 1988)) can be used to catalytically cleave mRNA transcripts to thereby inhibit translation of the protein encoded by the mRNA. A ribozyme having specificity for a CD40 or CD4OL
mRNA
can be designed based upon the nucleotide sequence of any of the CD40 or CD4OL
mRNA
sequences disclosed herein. For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in a CD40 or CD4OL mRNA (see, e.g., U.S.
Patent. Nos.
4,987,071 and 5,116,742). Alternatively, a CD40 or CD4OL mRNA can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA
molecules. See, e.g., Bartel et al., Science 261:1411-1418, 1993.
An inhibitory nucleic acid can also be a nucleic acid molecule that forms triple helical structures. For example, expression of a CD40 or CD4OL polypeptide can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the gene encoding the CD40 or CD4OL polypeptide (e.g., the promoter and/or enhancer, e.g., a sequence that is at least 1 kb, 2 kb, 3 kb, 4 kb, or 5 kb upstream of the transcription initiation start state) to form triple helical structures that prevent transcription of the gene in target cells. See generally Helene, Anticancer Drug Des. 6(6):569-84, 1991; Helene, Ann. N.Y.
Acad. Sci.
660:27-36, 1992; and Maher, Bioassays 14(12):807-15, 1992.

In various embodiments, inhibitory nucleic acids can be modified at the base moiety, sugar moiety, or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acids can be modified to generate peptide nucleic acids (see, e.g., Hyrup et al., Bioorg. Med.
Chem. 4(1):5-23, 1996). Peptide nucleic acids (PNAs) are nucleic acid mimics, e.g., DNA
mimics, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs allows for specific hybridization to DNA and RNA under conditions of low ionic strength.
The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols (see, e.g., Perry-O'Keefe et al., Proc. Natl. Acad. Sci.
U.S.A. 93:14670-675, 1996). PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication.
PNAs can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA
chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras can be generated which may combine the advantageous properties of PNA
and DNA. Such chimeras allow DNA recognition enzymes, e.g., RNAse H and DNA
polymerases, to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation.
The synthesis of PNA-DNA chimeras can be performed as described in Finn et al., Nucleic Acids Res. 24:3357-63, 1996. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry and modified nucleoside analogs. Compounds such as 5'-(4-methoxytrityl)amino-5'-deoxy-thymidine phosphoramidite can be used as a link between the PNA and the 5' end of DNA
(Mag et al., Nucleic Acids Res. 17:5973-88, 1989). PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5' PNA segment and a 3' DNA
segment (Finn et al., Nucleic Acids Res. 24:3357-63, 1996). Alternatively, chimeric molecules can be synthesized with a 5' DNA segment and a 3' PNA segment (Peterser et al., Bioorg. Med.
Chem. Lett. 5:1119-11124, 1975).

In some embodiments, the inhibitory nucleic acids can include other appended groups such as peptides, or agents facilitating transport across the cell membrane (see, Letsinger et al., Proc. Natl. Acad. Sci. U.S.A. 86:6553-6556, 1989; Lemaitre et al., Proc.
Natl. Acad. Sci.
U.S.A. 84:648-652, 1989; and WO 88/09810). In addition, the inhibitory nucleic acids can be modified with hybridization-triggered cleavage agents (see, e.g., Krol et al., Bio/Techniques 6:958-976, 1988) or intercalating agents (see, e.g., Zon, Pharm. Res. 5:539-549, 1988). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent, etc.
Another means by which expression of a CD40 or CD4OL mRNA can be decreased in a mammalian cell is by RNA interference (RNAi). RNAi is a process in which mRNA is degraded in host cells. To inhibit an mRNA, double-stranded RNA (dsRNA) corresponding to a portion of the gene to be silenced (e.g., a gene encoding a CD40 or CD4OL
polypeptide) is introduced into a mammalian cell. The dsRNA is digested into 21-23 nucleotide-long duplexes called short interfering RNAs (or siRNAs), which bind to a nuclease complex to form what is known as the RNA-induced silencing complex (or RISC). The RISC
targets the homologous transcript by base pairing interactions between one of the siRNA
strands and the endogenous mRNA. It then cleaves the mRNA about 12 nucleotides from the 3' terminus of the siRNA (see Sharp et al., Genes Dev. 15:485-490, 2001, and Hammond et al., Nature Rev.
Gen. 2:110-119, 2001).
RNA-mediated gene silencing can be induced in a mammalian cell in many ways, e.g., by enforcing endogenous expression of RNA hairpins (see, Paddison et al., Proc. Natl.
Acad. Sci. U.S.A. 99:1443-1448, 2002) or, as noted above, by transfection of small (21-23 nt) dsRNA (reviewed in Caplen, Trends Biotech. 20:49-51, 2002). Methods for modulating gene expression with RNAi are described, e.g., in U.S. Patent No. 6,506,559 and US
2003/0056235, which are hereby incorporated by reference.
Standard molecular biology techniques can be used to generate siRNAs. Short interfering RNAs can be chemically synthesized, recombinantly produced, e.g., by expressing RNA from a template DNA, such as a plasmid, or obtained from commercial vendors, such as Dharmacon. The RNA used to mediate RNAi can include synthetic or modified nucleotides, such as phosphorothioate nucleotides. Methods of transfecting cells with siRNA
or with plasmids engineered to make siRNA are routine in the art.

The siRNA molecules used to decrease expression of a CD40 or CD4OL mRNA can vary in a number of ways. For example, they can include a 3' hydroxyl group and strands of 21, 22, or 23 consecutive nucleotides. They can be blunt ended or include an overhanging end at either the 3' end, the 5' end, or both ends. For example, at least one strand of the RNA
molecule can have a 3' overhang from about 1 to about 6 nucleotides (e.g., 1-5, 1-3, 2-4, or 3-5 nucleotides (whether pyrimidine or purine nucleotides) in length. Where both strands include an overhang, the length of the overhangs may be the same or different for each strand.
To further enhance the stability of the RNA duplexes, the 3' overhangs can be stabilized against degradation (by, e.g., including purine nucleotides, such as adenosine or guanosine nucleotides or replacing pyrimidine nucleotides by modified analogues (e.g., substitution of uridine 2-nucleotide 3' overhangs by 2'-deoxythymidine is tolerated and does not affect the efficiency of RNAi). Any siRNA can be used in the methods of decreasing a CD40 or CD4OL mRNA, provided it has sufficient homology to the target of interest (e.g., a sequence present in any one of SEQ ID NOs: 56-61, e.g., a target sequence encompassing the translation start site or the first exon of the mRNA). There is no upper limit on the length of the siRNA that can be used (e.g., the siRNA can range from about 21 base pairs of the gene to the full length of the gene or more (e.g., about 20 to about 30 base pairs, about 50 to about 60 base pairs, about 60 to about 70 base pairs, about 70 to about 80 base pairs, about 80 to about 90 base pairs, or about 90 to about 100 base pairs).
Non-limiting examples of short interfering RNA (siRNA) that are CD40/CD4OL
inhibitors are described in, e.g., Pluvinet et al., Blood 104:3642-3646, 2004;
Karimi et al., Cell Immunol. 259(1):74-81, 2009; and Zheng et al., Arthritis Res. Ther.
12(1):R13, 2010.
Non-limiting examples of short hairpin RNA (shRNA) targeting CD40/CD4OL are described .. in Zhang et al., Gene Therapy 21:709-714, 2014. Non-limiting examples of microRNAs that are CD40/CD4OL inhibitors include, for example, miR146a (Chen et al., FEBS
Letters 585(3):567-573, 2011), miR-424, and miR-503 (Lee et al., Sci. Rep. 7:2528, 2017).
Non-limiting examples of aptamers that are CD40/CD4OL inhibitors are described in Soldevilla et al., Biomaterials 67:274-285, 2015.
In certain embodiments, a therapeutically effective amount of an inhibitory nucleic acid targeting a nucleic acid encoding a CD40 or CD4OL protein can be delivered locally to a subject (e.g., a human subject) in need thereof using any of the devices described herein.

In some embodiments, the inhibitory nucleic acid can be about 10 nucleotides to about 40 nucleotides (e.g., about 10 to about 30 nucleotides, about 10 to about 25 nucleotides, about 10 to about 20 nucleotides, about 10 to about 15 nucleotides, 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotides, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36 nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, or 40 nucleotides) in length. One skilled in the art will appreciate that inhibitory nucleic acids may comprise at least one modified nucleic acid at either the 5' or 3'end of DNA or RNA.
Any of the inhibitor nucleic acids described herein can be formulated for administration to the gastrointestinal tract. See, e.g., the formulation methods described in US 2016/0090598 and Schoellhammer et al., Gastroenterology, doi:
10.1053/j.gastro.2017.01.002, 2017.
As is known in the art, the term "thermal melting point (Tm)" refers to the temperature, under defined ionic strength, pH, and inhibitory nucleic acid concentration, at which 50% of the inhibitory nucleic acids complementary to the target sequence hybridize to the target sequence at equilibrium. In some embodiments, an inhibitory nucleic acid can bind specifically to a target nucleic acid under stringent conditions, e.g., those in which the salt concentration is at least about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30 C for short oligonucleotides (e.g., 10 to 50 nucleotide). Stringent conditions can also be achieved with the addition of destabilizing agents such as formamide.
In some embodiments of any of the inhibitory nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding CD40 or CD4OL) with a Tm of greater than 20 C, greater than 22 C, greater than 24 C, greater than 26 C, greater than 28 C, greater than 30 C, greater than 32 C, greater than 34 C, greater than 36 C, greater than 38 C, greater than 40 C, greater than 42 C, greater than 44 C, greater than 46 C, greater than 48 C, greater than 50 C, greater than 52 C, greater than 54 C, greater than 56 C, greater than 58 C, greater than 60 C, greater than 62 C, greater than 64 C, greater than 66 C, greater than 68 C, greater than 70 C, greater than 72 C, greater than 74 C, greater than 76 C, greater than 78 C, or greater than 80 C, e.g., as measured in phosphate buffered saline using a UV spectrophotometer.

In some embodiments of any of the inhibitor nucleic acids described herein, the inhibitory nucleic acid binds to a target nucleic acid (e.g., a nucleic acid encoding CD40 or CD4OL) with a Tm of about 20 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, about 30 C, about 28 C, about 26 C, about 24 C, or about 22 C
(inclusive); about 22 C
to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, about 30 C, about 28 C, about 26 C, or about 24 C (inclusive); about 24 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, about 30 C, about 28 C, or about 26 C (inclusive); about 26 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, about 30 C, or about 28 C
(inclusive); about 28 C
to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, about 32 C, or about 30 C
(inclusive); about 30 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, about 34 C, or about 32 C (inclusive); about 32 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, about 36 C, or about 34 C (inclusive); about 34 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, about 38 C, or about 36 C
(inclusive); about 36 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, about 40 C, or about 38 C (inclusive); about 38 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, about 42 C, or about 40 C
(inclusive); about 40 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, about 44 C, or about (inclusive); about 42 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, about 46 C, or about 44 C
(inclusive); about 44 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, about 48 C, or about 46 C (inclusive);
about 46 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, about 50 C, or about 48 C (inclusive); about 48 C
to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, about 52 C, or about 50 C (inclusive); about 50 C to about 80 C, about 78 C, about 76 C, about .. 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, about 54 C, or about 52 C (inclusive);
about 52 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, about 56 C, or about 54 C
(inclusive); about 54 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, about 58 C, or about 56 C (inclusive); about 56 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, about 60 C, or about 58 C (inclusive); about 58 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, about 62 C, or about 60 C (inclusive); about 60 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, about 64 C, or about 62 C (inclusive);
about 62 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, about 66 C, or about 64 C (inclusive); about 64 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, about 68 C, or about (inclusive); about 66 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, about 70 C, or about 68 C (inclusive); about 68 C to about 80 C, about 78 C, about 76 C, about 74 C, about 72 C, or about 70 C (inclusive); about 70 C to about 80 C, about 78 C, about 76 C, about 74 C, or about 72 C (inclusive); about 72 C to about 80 C, about 78 C, about 76 C, or about 74 C (inclusive); about 74 C to about 80 C, about 78 C, or about 76 C (inclusive); about 76 C to about 80 C or about 78 C
(inclusive); or about 78 C to about 80 C (inclusive).
In some embodiments, the inhibitory nucleic acid can be formulated in a nanoparticle (e.g., a nanoparticle including one or more synthetic polymers, e.g., Patil et al., Pharmaceutical Nanotechnol. 367:195-203, 2009; Yang et al., ACS Appl. Mater.
Interfaces, doi: 10.1021/acsami.6b16556, 2017; Perepelyuk et al., Mol. Ther. Nucleic Acids 6:259-268, 2017). In some embodiments, the nanoparticle can be a mucoadhesive particle (e.g., nanoparticles having a positively-charged exterior surface) (Andersen et al., Methods Mol.
Biol. 555:77-86, 2009). In some embodiments, the nanoparticle can have a neutrally-charged exterior surface.
In some embodiments, the inhibitory nucleic acid can be formulated, e.g., as a liposome (Buyens et al., I Control Release 158(3): 362-370, 2012; Scarabel et al., Expert Op/n. Drug Deliv. 17:1-14, 2017), a micelle (e.g., a mixed micelle) (Tangsangasaksri et al., BioMacromolecules 17:246-255, 2016; Wu et al., Nanotechnology, doi:
10.1088/1361-6528/aa6519, 2017), a microemulsion (WO 11/004395), a nanoemulsion, or a solid lipid nanoparticle (Sahay et al., Nature Biotechnol. 31:653-658, 2013; and Lin et al., Nanomedicine 9(1):105-120, 2014). Additional exemplary structural features of inhibitory nucleic acids and formulations of inhibitory nucleic acids are described in US
2016/0090598.
In some embodiments, a pharmaceutical composition can include a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In some examples, a pharmaceutical composition consists of a sterile saline solution and one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein). In certain embodiments, the sterile saline is a pharmaceutical grade saline. In certain embodiments, a pharmaceutical composition can include one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and sterile water. In certain embodiments, a pharmaceutical composition includes one or more inhibitory nucleic acid (e.g., any of the inhibitory nucleic acids described herein) and phosphate-buffered saline (PBS). In certain embodiments, a pharmaceutical composition consists of one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) and sterile phosphate-buffered saline (PBS). In some examples, the sterile saline is a pharmaceutical grade PBS.
In certain embodiments, one or more inhibitory nucleic acids (e.g., any of the inhibitory nucleic acids described herein) may be admixed with pharmaceutically acceptable active and/or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions depend on a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.
Pharmaceutical compositions including one or more inhibitory nucleic acids encompass any pharmaceutically acceptable salts, esters, or salts of such esters. Non-limiting examples of pharmaceutical compositions include pharmaceutically acceptable salts of inhibitory nucleic acids. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.
Also provided herein are prodrugs that can include additional nucleosides at one or both ends of an inhibitory nucleic acid which are cleaved by endogenous nucleases within the body, to form the active inhibitory nucleic acid.
Lipid moieties can be used to formulate an inhibitory nucleic acid. In certain such methods, the inhibitory nucleic acid is introduced into preformed liposomes or lipoplexes made of mixtures of cationic lipids and neutral lipids. In certain methods, inhibitory nucleic acid complexes with mono- or poly-cationic lipids are formed without the presence of a neutral lipid. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to a particular cell or tissue in a mammal. In some examples, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to fat tissue in a mammal. In certain embodiments, a lipid moiety is selected to increase distribution of an inhibitory nucleic acid to muscle tissue.
In certain embodiments, pharmaceutical compositions provided herein can include one or more inhibitory nucleic acid and one or more excipients. In certain such embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose, and polyvinylpyrrolidone.
In some examples, a pharmaceutical composition provided herein includes liposomes and emulsions. Liposomes and emulsions can be used to formulate hydrophobic compounds.
In some examples, certain organic solvents, such as dimethylsulfoxide, are used.
In some examples, a pharmaceutical composition provided herein includes one or more tissue-specific delivery molecules designed to deliver one or more inhibitory nucleic acids to specific tissues or cell types in a mammal. For example, a pharmaceutical composition can include liposomes coated with a tissue-specific antibody.
In some embodiments, a pharmaceutical composition provided herein can include a co-solvent system. Examples of such co-solvent systems include benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. A non-limiting example of such a co-solvent system is the VPD co-solvent system, which is a solution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80 and 65% w/v polyethylene glycol 300. As can be appreciated, other surfactants may be used instead of Polysorbate 80TM; the fraction size of polyethylene glycol may be varied;
other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone;
and other sugars or polysaccharides may substitute for dextrose. Any of the pharmaceutical compositions described herein can be delivered locally to a subject using any of the devices described herein.
In some examples, an inhibitory nucleic acid can be formulated to include a carrier and is formulated in aqueous solution, such as water or physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer.
In some examples, other ingredients are included (e.g., ingredients that aid in solubility or serve as preservatives). In some examples, an inhibitory nucleic acid can be formulated as a suspension and can be prepared using appropriate liquid carriers, suspending agents, and the like. An inhibitory nucleic acid can be formulated as a suspension, solution, or emulsion in oily or aqueous vehicles prior to intrathecal administration using any of the devices described herein, and may contain formulatory agents such as suspending, stabilizing, and/or dispersing agents. Solvents suitable for formulating an inhibitory nucleic acid include, but are not limited to, lipophilic solvents and fatty oils, such as sesame oil, synthetic fatty acid esters, such as ethyl oleate or triglycerides, and liposomes.
Antibodies In some embodiments, the CD40/CD4OL inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, an antibody or antigen-binding fragment described herein binds specifically to CD40 or CD4OL, or to both CD40 and CD4OL.
In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv)2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat.
Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC
(Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO
Mol. Med.
9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, la-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG
(L,H)-Fc, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Patent No. 5,759,808; Stijlemans et al., I Biol.
Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., 1 Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol.
25(2):85-91, 2004), Diabody-CH3 (Guo et al.õ Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab')2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther.
8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.
Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab')2 fragment, and a Fab' fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgGl, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgGl, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgAl or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgAl or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).
In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986;
WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., I Exp. Med.
175:217-225, 1992; Kolstelny et al., I Immunol. 148(5):1547-1553, 1992; Hollinger et al., Proc. Natl.
Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., I Immunol. 152:5368, 1994; Tutt et al., 1 Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC
Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (Fab' scFv)2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Patent No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., I Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.
In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Patent Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895;
US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US
2017/0002082, each of which is incorporated by reference in its entirety.
In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of PG102 (Pangenetics) (Bankert et al., I Immunol.
194(9):4319-4327, 2015); 2C10 (Lowe et al., Am. I Transplant 12(8):2079-2087, 2012); ASKP1240 (Bleselumab) (Watanabe et al., Am. I Transplant 13(8):1976-1988, 2013); 4D11 (Imai et al., Transplantation 84(8):1020-1028, 2007); BI 655064 (Boehringer Ingelheim) (Visvanathan et al., 2016 American College of Rheumatology Annual Meeting, Abstract 1588, September 28, 2016); 5D12 (Kasran et al., Aliment. Pharmacol. Ther., 22(2):111-122, 2005;
Boon et al., Toxicology 174(1):53-65, 2002); ruplizumab (hu5c8) (Kirk et al., Nat. Med.
5(6):686-693, 1999); CHIR12.12 (HCD122) (Weng et al., Blood 104(11):3279, 2004; Tai et al., Cancer Res.
65(13):5898-5906, 2005); CDP7657 (Shock et al., Arthritis Res. Ther.
17(1):234, 2015);
BMS-986004 domain antibody (dAb) (Kim et al., Am. I Transplant. 17(5):1182-1192, 2017);
5c8 (Xie et al., I Immunol. 192(9):4083-4092, 2014); dacetuzumab (SGN-40) (Lewis et al., Leukemia 25(6):1007-1016, 2011; and Khubchandani et al., Curr. Opin. Investig.
Drugs 10(6):579-587, 2009); lucatumumab (HCD122) (Bensinger et al., Br. I Haematol.
159: 58-66, 2012; and Byrd et al., Leuk. Lymphoma 53(11):
10.3109/10428194.2012.681655, 2012);
PG102 (FFP104) (Bankert et al., I Immunol. 194(9):4319-4327, 2015); Chi Lob (Johnson et al., I Clin. Oncol. 28:2507, 2019); and ASKP1240 (Okimura et al., Am.
Transplant. 14(6): 1290-1299, 2014; or Ma et al., Transplantation 97(4): 397-404, 2014).
Further teachings of CD40/CD4OL antibodies and antigen-binding fragments thereof are described in, for example, U.S. Patent Nos. 5,874,082; 7,169,389;
7,271,152; 7,288,252;
7,445,780; 7,537,763, 8,277,810; 8,293,237, 8,551,485; 8,591,900; 8,647,625;
8,784,823;

8,852,597; 8,961,976; 9,023,360, 9,028,826; 9,090,696, 9,221,913;
US2014/0093497; and US2015/0017155 each of which is incorporated by reference in its entirety.
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (KD) of less than 1 x 10-5M (e.g., less than 0.5 x 10-5M, less than 1 x 10' M, less than 0.5 x 10' M, less than 1 x 10-7M, less than 0.5 x 10-7M, less than 1 x 10-8M, less than 0.5 x 10-8M, less than 1 x 10-9M, less than 0.5 x 10-9M, less than 1 x 10-10 M, less than 0.5 x 10-10 M, less than 1 x 10-11M, less than 0.5 x 10-11M, or less than 1 x 10-12M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a KID of about 1 x 10-12M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8 M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-1 M, about 0.5 x 10-10 M, about 1 x 10-11M, or about 0.5 x 10-11M (inclusive); about 0.5 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9 M, about 1 x 1010 M, about 0.5 x 10-1 M, or about 1 x 10-11M (inclusive); about 1 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 1010 M, or about 0.5 x 10-10 M (inclusive); about 0.5 x 1010 M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, or about 1 x 10-10 M
(inclusive);
about 1 x 1010 M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9 M, or about 0.5 x 10-9M (inclusive); about 0.5 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, or about 1 x 10-9M (inclusive); about 1 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, about 1 x 10-8M, or about 0.5 x 10-8M (inclusive); about 0.5 x 10-8M to about 1 x 10-5 M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, or about 1 x 10-8M (inclusive); about 1 x 10-8M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10-6M, about 1 x 10-7M, or about 0.5 x 10-7M
(inclusive);
about 0.5 x 10-7M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, or about 1 x 10-7 M (inclusive); about 1 x 10-7 M to about 1 x 10-5 M, about 0.5 x 10-5 M, about 1 x 10-6 M, or about 0.5 x 10-6 M (inclusive); about 0.5 x 10-6 M to about 1 x 10-5 M, about 0.5 x 10-5 M, or about 1 x 10-6 M (inclusive); about 1 x 10-6 M to about 1 x 10-5 M or about 0.5 x 10-5 M (inclusive); or about 0.5 x 10-5 M to about 1 x 10-5 M
(inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Kar of about lx 10-6 s-1 to about lx 10-3 s-1, about 0.5 x 10-3 s-1, about lx 10-4 s-1, about 0.5 x 10-4 s-1, about 1 x 10-5 s-1, or about 0.5 x 10-5 s-1 (inclusive); about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, about 0.5 x 10-4 s-1, or about 1 x 10-5 s-1 (inclusive); about 1 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, or about 0.5 x 10-4 s-1 (inclusive); about 0.5 x 10-4 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, or about 1 x 10-4 s-1 (inclusive); about 1 x 10-4 s-1 to about 1 x 10-3 s-1, or about 0.5 x 10-3 s-1 (inclusive); or about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Kon of about 1 x 102 M's' to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105 M's', about 0.5 x 105 M's', about 1 x 104 N4-1S-1, about 0.5 x 104 M's', about 1 x 103 M's', or about 0.5 x 103 M-1s-1 (inclusive); about 0.5 x 103 M-1s-1 to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105 M's', about 0.5 x 105 M's', about 1 x 104 M's', about 0.5 x iO4 M's', or about 1 x 103 M-1s-1 (inclusive); about 1 x 103 M-1s-1 to about 1 x 106 M-1s-1, about 0.5 x 106 m-ls-1, about 1 x 105 M's', about 0.5 x 105 M's', about 1 x 104 M's', or or about 0.5 x 104 (inclusive); about 0.5 x 104 ws-1 to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105 M's', about 0.5 x 105 M's', or about 1 x 104 M-1s-1 (inclusive);
about 1 x 104 M's' to about 1 x 106 M's', about 0.5 x 106 m-ls-1, about 1 x 105 M's', or about 0.5 x 105 M-1s-1 (inclusive); about 0.5 x 105 M-1s-1 to about 1 x 106 M's', about 0.5 x 106 M's', or about 1 x 105 M-1s-1 (inclusive); about 1 x 105 M-1s-1 to about 1 x 106 M's', or about 0.5 x 106 m-ls-1 (inclusive); or about 0.5 x 106 ws-1 to about 1 x 106 M-1s-1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
Fusion and Truncated Proteins and Peptides In some embodiments, the CD40/CD4OL inhibitor is a fusion protein, a truncated protein (e.g., a soluble receptor) or a peptide. In some embodiments, the inhibitor is a truncated protein as disclosed in, for example, WO 01/096397.
In some embodiments, the CD40/CD4OL inhibitor is a peptide, such as a cyclic peptide (see, e.g., U.S.
Patent No. 8,802,634; Bianco et al., Org. Biomol. Chem. 4:1461-1463, 2006;
Deambrosis et al., I Mot. Med. 87(2):181-197, 2009; Vaitaitis et al., Diabetologia 57(11):2366-2373, 2014).
In some embodiments, the CD40/CD4OL inhibitor is a CD40 ligand binder, for example, a Tumor Necrosis Factor Receptor-associated Factor (TRAF): TRAF2, TRAF3, TRAF6, TRAF5 and TTRAP, or E3 ubiquitin-protein ligase RNF128.
Small Molecules In some embodiments, the CD40/CD4OL inhibitor is a small molecule (see, e.g., U.S.
Patent No. 7,173,046, U.S. Patent Application No. 2011/0065675). In some embodiments, the small molecule is Bio8898 (Silvian et al., ACS Chem. Biol. 6(6):636-647, 2011); Suramin (Margolles-Clark et al., Biochem. Pharmacol. 77(7):1236-1245, 2009); a small-molecule organic dye (Margolles-Clark et al., I Mot. Med. 87(11):1133-1143, 2009;
Buchwald et al., Mot. Recognit. 23(1):65-73, 2010), a naphthalenesulphonic acid derivative (Margolles-Clark et al., Chem. Biol. Drug Des. 76(4):305-313, 2010), or a variant thereof.
CD3 Inhibitors The term "CD3 inhibitor" refers to an agent which decreases the ability of one or more of CD3-y, CD3o, CD3c, and CD3t to associate with one or more of TCR-a, TCR-I3, TCR-6, and TCR-7. In some embodiments, the CD3 inhibitor can decrease the association between one or more of CD37, CD3o, CD3c, and CD3t and one or more of TCR-a, TCR-I3, TCR-6, and TCR-7 by blocking the ability of one or more of CD37, CD3o, CD3c, and CD3 to interact with one or more of TCR-a, TCR-I3, TCR-6, and TCR-7.
In some embodiments, the CD3 inhibitor is an antibody or an antigen-binding fragment thereof, a fusion protein, or a small molecule. Exemplary CD3 inhibitors are described herein. Additional examples of CD3 inhibitors are known in the art.
Exemplary sequences for human CD37, human CD3o, human CD3c, and human CD3t are shown below.
Human CD3y (SEQ ID NO: 62) meqgkglavl ilaiillqgt laqsikgnhl vkvydyqedg svlltcdaea knitwfkdgk migfltedkk kwnlgsnakd prgmyqckgs qnkskplqvy yrmcqnciel naatisgflf aeivsifvla vgvyfiagqd gvrqsrasdk qtllpndqly qplkdreddq yshlqgnqlr rn Human CD38 Isoform A (SEQ ID NO: 63) fkipieele drvfvncnts itwvegtvgt llsditrldl gkrildprgi yrcngtdiyk dkestvqvhy rmcqscveld patvagiivt dviatlllal gvfcfaghet grlsgaadtq allmdqvyq plrdrddaqy shlggnwarn k Human CD38 Isoform B (SEQ ID NO: 64) fkipieele drvfvncnts itwvegtvgt llsditrldl gkrildprgi yrcngtdiyk dkestvqvhy rtadtqallr ndqvyqp1rd rddaqyshlg gnwarnk Human CD3 c (SEQ ID NO: 65) dgneemgg itqtpykvsi sgttviltcp qypgseilwq hndkniggde ddknigsded hlslkefsel eqsgyyvcyp rgskpedanf ylylrarvce ncmemdvmsv ativivdici tgg1111vyy wsknrkakak pvtrgagagg rqrgqnkerp ppvpnpdyep irkgqrdlys glnqrri Human CD3 4 Isoform 1 (SEQ ID NO: 66) qsfglldpk lcylldgilf iygviltalf lrvkfsrsad apayqqgqnq lynelnlgrr eeydvldkrr grdpemggkp qrrknpqegl ynelqkdkma eayseigmkg errrgkghdg lyqglstatk dtydalhmqa 1ppr Human CD3 4 Isoform 2 (SEQ ID NO: 67) qsfglldpk lcylldgilf iygviltalf lrvkfsrsad apayqqgqnq lynelnlgrr eeydvldkrr grdpemggkp rrknpqegly nelqkdkmae ayseigmkge rrrgkghdgl yqglstatkd tydalhmqal ppr Antibodies In some embodiments, the CD3 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, the CD3 inhibitor is an antibody or antigen-binding fragment that binds specifically to CD3-y. In some embodiments, the CD3 inhibitor is an antibody or antigen-binding fragment that binds specifically to CD3o.
In some mebodiments, the CD3 inhibitor is an antibody or antigen-binding fragment that binds specifically to CD3 E. In some embodiments, the CD3 inhibitor is an antibody or antigen-binding fragment that binds specifically to CDK In some embodiments, the CD3 inhibitor is an antibody or an antigen-binding fragment that can bind to two or more (e.g., two, three, or four) of CD37, CD3o, CD3c, and CD3.
In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv)2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat.
Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mot. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC
(Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO
Mot. Med.
9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, la-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG
(L,H)-Fc, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Patent No. 5,759,808; Stijlemans et al., I Biol.
Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al.,1 Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol.
25(2):85-91, 2004), Diabody-CH3 (Guo et al.õ Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab')2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther.
8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.
Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab')2 fragment, and a Fab' fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgGl, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgGl, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgAl or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgAl or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).
In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986;
WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby etal., I Exp. Med.
175:217-225, 1992; Kolstelny etal., I Immunol. 148(5):1547-1553, 1992; Hollinger etal., Proc. Natl.
Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., I Immunol. 152:5368, 1994; Tutt et al., 1 Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC
Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (Fab' scFv)2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt etal., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Patent No. 4,676,980), a linear antibody (Zapata etal., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., I Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.
In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Patent Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895;
US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US
2017/0002082, each of which is incorporated by reference in its entirety.
In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of visiluzumab (Nuvion; HuM-291; M291; SMART anti-CD3 antibody) (Carpenter et al., Biol. Blood Marrow Transplant 11(6): 465-471, 2005;
Trajkovic Curr. Opin.

Investig. Drugs 3(3): 411-414, 2002; Malviya etal., I Nucl. Med. 50(10): 1683-1691, 2009);
muromonab-CD3 (orthoclone OKT3) (Hon i etal., Surg. Today 41(4): 585-590, 2011; Norman Ther. Drug Monit. 17(6): 615-620, 1995; and Gramatzki et al., Leukemia 9(3):
382-390, 19);
otelixizumab (TRX4) (Vossenkamper et al., Gastroenterology 147(1): 172-183, 2014; and Wiczling et al., I Cl/n. Pharmacol. 50(5): 494-506, 2010); foralumab (NI-0401) (Ogura et al., Cl/n. Immunol. 183: 240-246; and van der Woude etal., Inflamm. Bowel Dis.
16: 1708-1716, 2010); ChAgly CD3; teplizumab (MGA031) (Waldron-Lynch et al., Sci.
Transl. Med.
4(118): 118ra12, 2012; and Skelley et al., Ann. Pharmacother 46(10): 1405-1412, 2012); or catumaxomab (Removabg) (Linke et al., Mabs 2(2): 129-136, 2010; and Bokemeyer et al., Gastric Cancer 18(4): 833-842, 2015).
Additional examples of CD3 inhibitors that are antibodies or antibody fragments are described in, e.g., U.S. Patent Application Publication Nos. 2017/0204194, 2017/0137519, 2016/0368988, 2016/0333095, 2016/0194399, 2016/0168247, 2015/0166661, 2015/0118252, 2014/0193399, 2014/0099318, 2014/0088295, 2014/0080147, 2013/0115213, 2013/0078238, 2012/0269826, 2011/0217790, 2010/0209437, 2010/0183554, 2008/0025975, 2007/0190045, 2007/0190052, 2007/0154477, 2007/0134241, 2007/0065437, 2006/0275292, 2006/0269547, 2006/0233787, 2006/0177896, 2006/0165693, 2006/0088526, 2004/0253237, 2004/0202657, 2004/0052783, 2003/0216551, and 2002/0142000, each of which is herein incorporated by reference in its entirety (e.g., the sections describing the CD3 inhibitors).
Additional CD3 inhibitors that are antibodies or antigen-binding antibody fragments are described in, e.g., Smith etal., I Exp. Med. 185(8):1413-1422, 1997; Chatenaud etal., Nature 7:622-632, 2007.
In some embodiments, the CD3 inhibitor comprises or consists of a bispecific antibody (e.g., JNJ-63709178) (Gaudet et al., Blood 128(22): 2824, 2016); JNJ-(Girgis etal., Blood 128: 5668, 2016); MGD009 (Tolcher etal., I Cl/n. Oncol.
34:15, 2016);
.. ERY974 (Ishiguro et al., Sci. Transl. Med. 9(410): pii.eaa14291, 2017);
A1V1V564 (Hoseini and Cheung Blood Cancer 1 7:e522, 2017); AFM11 (Reusch et al.MAbs 7(3): 584-604, 2015); duvortuxizumab (JNJ 64052781); R06958688; blinatumomab (Blincytog;
AMG103) (Ribera Expert Rev. Hematol. 1:1-11, 2017; and Mori etal., N Engl. I Med.
376(23):e49, 2017); XmAb13676; or REGN1979 (Bannerji et al., Blood 128: 621, 2016; and Smith et al., Sci. Rep. 5:17943, 2015)).
In some embodiments, the CD3 inhibitor comprises or consists of a trispecific antibody (e.g., ertumaxomab (Kiewe and Thiel, Expert Op/n. Investig. Drugs 17(10): 1553-1558, 2008; and Haense et al., BMC Cancer 16:420, 2016); or FBTA05 (Bi20;
Lymphomun) (Buhmann etal., I Transl. Med. 11:160, 2013; and Schuster etal., Br. I
Haematol. 169(1):
90-102, 2015)).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (KD) of less than 1 x 10-5M (e.g., less than 0.5 x 10-5M, less than 1 x 10' M, less than 0.5 x 10' M, less than 1 x 10-7M, less than 0.5 x 10-7M, less than 1 x 10-8M, less than 0.5 x 10-8M, less than 1 x 10-9M, less than 0.5 x 10-9M, less than 1 x 10-10 M, less than 0.5 x 10-10 M, less than 1 x 10-11M, less than 0.5 x 10-11M, or less than 1 x 10-12M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a KID of about 1 x 10-12M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8 M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-1 M, about 0.5 x 10-10 M, about 1 x 10-11M, or about 0.5 x 10-11M (inclusive); about 0.5 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9 M, about 1 x 1010 M, about 0.5 x 10-1 M, or about 1 x 10-11M (inclusive); about 1 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 1010 M, or about 0.5 x 10-10 M (inclusive); about 0.5 x 1010 M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, or about 1 x 10-10 M
(inclusive);
about 1 x 1010 M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9 M, or about 0.5 x 10-9M (inclusive); about 0.5 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, or about 1 x 10-9M (inclusive); about 1 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, about 1 x 10-8M, or about 0.5 x 10-8M (inclusive); about 0.5 x 10-8M to about 1 x 10-5 M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, or about 1 x 10-8M (inclusive); about 1 x 10-8M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10-6M, about 1 x 10-7M, or about 0.5 x 10-7M
(inclusive);
about 0.5 x 10-7M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, or about 1 x 10-7M (inclusive); about 1 x 10-7M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10-6M, or about 0.5 x 10-6M (inclusive); about 0.5 x 10-6M to about 1 x 10-5M, about 0.5 x 10-5M, or about 1 x 10-6M (inclusive); about 1 x 10-6M to about 1 x 10-5M or about 0.5 x 10-5M (inclusive); or about 0.5 x 10-5M to about 1 x 10-5M
(inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Kar of about lx 10-6 s-1 to about lx 10-3 s-1, about 0.5 x 10-3 s-1, about lx 10-4 s-1, about 0.5 x 10-4 s-1, about 1 x 10-5 s-1, or about 0.5 x 10-5 s-1 (inclusive); about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, about 0.5 x 10-4 s-1, or about 1 x 10-5 s-1 (inclusive); about 1 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, or about 0.5 x 10-4 s-1 (inclusive); about 0.5 x 10-4 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, or about 1 x 10-4 s-1 (inclusive); about 1 x 10-4 s-1 to about 1 x 10-3 s-1, or about 0.5 x 10-3 s-1 (inclusive); or about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Kon of about 1 x 102 M's' to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105M-1s-1, about 0.5 x 105M-1s-1, about 1 x 104 N4-1S-1, about 0.5 x 104M-1s-1, about 1 x 103 M's', or about 0.5 x iO3 M's' (inclusive); about 0.5 x 103M-1s-1-to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x i05 M's', about 0.5 x 10 M's', about 1 x iO4 M's', about 0.5 x iO4 M's', or about 1 x iO3 M's' (inclusive); about 1 x iO3 M's' to about 1 x 106 M-1s-1, about 0.5 x 106 m-ls-1, about 1 x 10 M's', about 0.5 x 10 M's', about 1 x 104M-1s-1, or about 0.5 x 104 (inclusive); about 0.5 x 104 ws-1 to about 1 X 106 M's', about 0.5 x 106 M's', about 1 x i05 M's', about 0.5 x i05 M's', or about 1 x iO4 M's' (inclusive);
about 1 x 104N4-1s --1 to about 1 x 106 M's', about 0.5 X 106 m-ls-1, about 1 X 105 M's', or about 0.5 x i05 M's' (inclusive); about 0.5 x 10 M's' to about 1 x 106 M's', about 0.5 x s, or about 1 x 105M-1s-1 (inclusive); about 1 x 10 M's' to about 1 x 106 M's', or about 0.5 x 106 m-ls-1 (inclusive); or about 0.5 x 106 ws-1 to about 1 X 106 M-1s-1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
Fusion and Truncated Proteins and Peptides In some embodiments, the CD3 inhibitor is a fusion protein, a truncated protein (e.g., a soluble receptor), or a peptide. In some embodiments, the CD3 inhibitor can be a fusion protein (see, e.g., Lee et al., Oncol. Rep. 15(5): 1211-1216, 2006).

Small Molecules In some embodiments, the CD3 inhibitor comprises or consists of a bispecific small molecule-antibody conjugate (see, e.g., Kim et al., PNAS 110(44): 17796-17801, 2013; Viola et al., Eur. I Immunol. 27(11):3080-3083, 1997).
CD14 Inhibitors The term "CD14 inhibitors" refers to an agent which decreases the ability of CD14 to bind to lipopolysaccharide (LPS). CD14 acts as a co-receptor with Toll-like receptor 4 (TLR4) that binds LPS in the presence of lipopolysaccharide-binding protein (LBP).
In some embodiments, the CD14 inhibitor can decrease the binding between CD14 and LPS
by blocking the ability of CD14 to interact with LPS.
In some embodiments, the CD14 inhibitor is an antibody or an antigen-binding fragment thereof In some embodiments, the CD14 inhibitor is a small molecule.
Exemplary CD14 inhibitors are described herein. Additional examples of CD14 inhibitors are known in the art.
An exemplary sequence for human CD14 is shown below.
Human CD14 (SEQ ID NO: 68) maaaaasrgv gaklglreir ihlcqrspgs qgvrdfiekr yvelkkanpd 1pilirecsd vqpklwarya fgqetnvpin nfsadqvtralenvlsgka CD14 Inhibitors- Antibodies In some embodiments, the CD14 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, the CD14 inhibitor is an antibody or antigen-binding fragment that binds specifically to CD14.
In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv)2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat.
Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC
(Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula etal., EMBO Mol.
Med.
9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume etal., Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, la-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG
(L,H)-Fc, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U .S . Patent No. 5,759,808; Stijlemans et al., I Biol.
Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., I Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol.
25(2):85-91, 2004), Diabody-CH3 (Guo et al.õ Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab')2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston etal., Human Antibodies 10(3-4):127-142, 2001; Wheeler etal., Mol. Ther.
8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.
Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab')2 fragment, and a Fab' fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgGl, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgGl, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgAl or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgAl or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).
In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986;
WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby etal., I Exp. Med.
175:217-225, 1992; Kolstelny etal., I Immunol. 148(5):1547-1553, 1992; Hollinger etal., Proc. Natl.
Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., I Immunol. 152:5368, 1994; Tutt et al., Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC
Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (Fab' scFv)2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt etal., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Patent No. 4,676,980), a linear antibody (Zapata etal., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., I Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.
In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Patent Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895;
US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US
2017/0002082, each of which is incorporated by reference in its entirety.
In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of IC14 (Axtelle and Pribble, I Endotoxin Res. 7(4): 310-314, 2001;
Reinhart etal., Crit. Care Med. 32(5): 1100-1108, 2004; Spek etal., I Clin.
Immunol. 23(2):
132-140, 2003). Additional examples of anti-CD14 antibodies and CD14 inhibitors can be found, e.g., in WO 2015/140591 and WO 2014/122660, incorporated in its entirety herein.

Additional examples of CD14 inhibitors that are antibodies or antibody fragments are described in, e.g., U.S. Patent Application Serial No. 2017/0107294, 2014/0050727, 2012/0227412, 2009/0203052, 2009/0029396, 2008/0286290, 2007/0106067, 2006/0257411, 2006/0073145, 2006/0068445, 2004/0092712, 2004/0091478, and 2002/0150882, each of which is herein incorporated by reference (e.g., the sections that describe CD14 inhibitors).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (KD) of less than 1 x 10-5M (e.g., less than 0.5 x 10-5M, less than 1 x 10' M, less than 0.5 x 10' M, less than 1 x 10-7M, less than 0.5 x 10-7M, less than 1 x 10-8M, less than 0.5 x 10-8M, less than 1 x 10-9M, less than 0.5 x 10-9M, less than 1 x 10-10 M, less than 0.5 x 10-10 M, less than 1 x 10-11M, less than 0.5 x 10-11M, or less than 1 x 10-12M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a KID of about 1 x 10-12M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8 M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-10 M, about 0.5 x 10-10 M, about 1 x 10-11M, or about 0.5 x 10-11M (inclusive); about 0.5 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9 M, about 1 x 10-10 M, about 0.5 x 10-1 M, or about 1 x 10-11M (inclusive); about 1 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-10 M, or about 0.5 x 10-10 M (inclusive); about 0.5 x 10-10 M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, or about 1 x 10-10 M
(inclusive);
about 1 x 10-10 M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9 M, or about 0.5 x 10-9M (inclusive); about 0.5 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, or about 1 x 10-9M (inclusive); about 1 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, about 1 x 10-8M, or about 0.5 x 10-8M (inclusive); about 0.5 x 10-8M to about 1 x 10-5 M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, or about 1 x 10-8 M (inclusive); about 1 x 10-8 M to about 1 x 10-5 M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10' M, or about 0.5 x 10' M
(inclusive);
about 0.5 x 10' M to about 1 x 10-5 M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, or about 1 x 10' M (inclusive); about 1 x 10' M to about 1 x 10-5 M, about 0.5 x 10-5 M, about 1 x 10' M, or about 0.5 x 10' M (inclusive); about 0.5 x 10' M to about 1 x 10-5 M, about 0.5 x 10-5 M, or about 1 x 10' M (inclusive); about 1 x 10' M to about 1 x 10-5 M or about 0.5 x 10-5 M (inclusive); or about 0.5 x 10-5 M to about 1 x 10-5 M
(inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Kar of about lx 10' s-1 to about lx 10-3 s-1, about 0.5 x 10-3 s-1, about lx 10-4 s-1, about 0.5 x 10-4 s-1, about 1 x 10-5 s-1, or about 0.5 x 10-5 s-1 (inclusive); about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, about 0.5 x 10-4 s-1, or about 1 x 10-5 s-1 (inclusive); about 1 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, or about 0.5 x 10-4 s-1 (inclusive); about 0.5 x 10-4 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, or about 1 x 10-4 s-1 (inclusive); about 1 x 10-4 s-1 to about 1 x 10-3 s-1, or about 0.5 x 10-3 s-1 (inclusive); or about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Kon of about 1 x 102 M's' to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105 M's', about 0.5 x 105 M's', about 1 x 104 N4-1S-1, about 0.5 x 104 M's', about 1 x 103 M's', or about 0.5 x 103 M-1s-1 (inclusive); about 0.5 x 103 M-1s-1 to about 1 x 106 N4-1s-1, about 0.5 x 106 M's', about 1 x 105 M's', about 0.5 x 105 M's', about 1 x 104 M's', about 0.5 x 104 M's', or about 1 x 103 M-1s-1 (inclusive); about 1 x 103 M-1s-1 to about 1 x 106 M-1s-1, about 0.5 x 106 m-ls-1, about 1 x 105 M's', about 0.5 x 105 M's', about 1 x 104 M's', or about 0.5 x 104 N4-1s-1 (inclusive); about 0.5 x 104 ws-1 to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105 M's', about 0.5 x 105 M's', or about 1 x 104 M-1s-1 (inclusive);
about 1 x 104 M's' to about 1 x 106 M's', about 0.5 x 106 m-ls-1, about 1 x 105 M's', or about 0.5 x 105 M-1s-1 (inclusive); about 0.5 x 105 M-1s-1 to about 1 x 106 M's', about 0.5 x 106 M's', or about 1 x 105 M-1s-1 (inclusive); about 1 x 105 M-1s-1 to about 1 x 106 M's', or .. about 0.5 x 106 m-ls-1 (inclusive); or about 0.5 x 106 ws-1 to about 1 x 106 M-1s-1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
Additional examples of CD14 inhibitors that are antibodies or antigen-binding fragments are known in the art.

CD14 Inhibitors- Small Molecules In some embodiments, the CD14 inhibitor is a small molecule. Non-limiting examples of CD14 inhibitors that are small molecules are described in, e.g., methyl 6-deoxy-6-N-dimethyl-N-cyclopentylammonium-2, 3-di-O-tetradecyl-a-D-glucopyranoside iodide (IAXO-101); methyl 6-Deoxy-6-amino-2,3-di-O-tetradecyl-a-D-glucopyranoside (IAX0-102); N-(3,4-bis-tetradecyloxy-benzy1)-N-cyclopentyl-N,N-dimethylammonium iodide (IAXO-103); and IMO-9200.
Additional examples of CD14 inhibitors that are small molecules are known in the art.
CD20 Inhibitors The term "CD20 inhibitors" refers to an agent that binds specifically to CD20 expressed on the surface of a mammalian cell.
In some embodiments, the CD20 inhibitor is an antibody or an antigen-binding fragment thereof, or a fusion protein or peptide. Exemplary CD20 inhibitors are described herein. Additional examples of CD20 inhibitors are known in the art.
An exemplary sequence of human CD20 is shown below.
Human CD20 (SEQ ID NO: 69) mttprnsvng tfpaepmkgp iamqsgpkpl frrmsslvgp tqsffmresk tlgavqimng lfhialggll mipagiyapi cvtvwyplwg gimyiisgsl laateknsrk clvkgkmimn slslfaaisg milsimdiln ikishflkme slnfirahtp yiniyncepa npseknspst qycysiqslf lgilsvmlif affqelviag ivenewkrtc srpksnivll saeekkeqti eikeevvglt etssqpknee dieiipiqee eeeetetnfp eppqdqessp iendssp CD20 Inhibitors- Antibodies In some embodiments, the CD20 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv).
In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv)2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat.
Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC
(Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula etal., EMBO Mol.
Med.
9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume etal., Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, la-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG
(L,H)-Fc, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Patent No. 5,759,808; Stijlemans et al., I Biol.
Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al.,1 Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol.
25(2):85-91, 2004), Diabody-CH3 (Guo et al.õ Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab')2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston etal., Human Antibodies 10(3-4):127-142, 2001; Wheeler etal., Mol. Ther.
8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.
Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab')2 fragment, and a Fab' fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgGl, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgGl, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgAl or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgAl or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).
In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986;
WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby etal., I Exp. Med.
175:217-225, 1992; Kolstelny etal., I Immunol. 148(5):1547-1553, 1992; Hollinger etal., Proc. Natl.
Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., I Immunol. 152:5368, 1994; Tutt et al., Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC
Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (Fab' scFv)2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt etal., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Patent No. 4,676,980), a linear antibody (Zapata etal., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., I Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.
In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Patent Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895;
US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US
2017/0002082, each of which is incorporated by reference in its entirety.
In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of rituximab (Rituxang, MabTherag, MK-8808) (Ji et al., Indian Hematol. Blood Transfus. 33(4): 525-533, 2017; and Calderon-Gomez and Panes Gastroenterology 142(1): 1741-76, 2012); - PF-05280586; ocrelizumab (OcrevusTM) (Sharp N. Engl. I Med. 376(17): 1692, 2017); ofatumumab (Arzerrag; HuMax-CD20) (AlDallal Ther. Clin. Risk Manag. 13:905-907, 2017; and Furman etal., Lancet Haematol.
4(1): e24-e34, 2017); PF-05280586 (Williams et al., Br. I Cl/n. Pharmacol. 82(6): 1568-1579, 2016;
and Cohen et al., Br. I Cl/n. Pharmacol. 82(1): 129-138, 2016); obinutuzumab (Gazyvag) (Reddy et al., Rheumatology 56(7): 1227-1237, 2017; and Marcus et al., N.
Engl. I Med.
377(14): 1331-1344, 2017); ocaratuzumab (AME-133v; LY2469298) (Cheney et al., Mabs 6(3): 749-755, 2014; and Tobinai et al., Cancer Sci. 102(2): 432-8, 2011);
GP2013 (Jurczak et al., Lancet Haenatol. 4(8): e350-e361, 2017); IBI301; HLX01; veltuzumab (hA20) (Kalaycio et al., Leuk. Lymphoma 57(4): 803-811, 2016; and Ellebrecht et al., JAMA
Dermatol. 150(12): 1331-1335, 2014); SCT400 (Gui et al., Chin. I Cancer Res.
28(2): 197-208); ibritumomab tiuxetan (Zevaling) (Philippe et al., Bone Marrow Transplant 51(8):
1140-1142, 2016; and Lossos et al., Leuk. Lymphoma 56(6): 1750-1755, 2015);
ublituximab (TG1101) (Sharman et al., Blood 124: 4679, 2014; and Sawas et al., Br. I
Haematol. 177(2):
243-253, 2017); LFB-R603 (Esteves et al., Blood 118: 1660, 2011; and Baritaki et al., Int.
Oncol. 38(6): 1683-1694, 2011); or tositumomab (Bexxar) (Buchegger et al., I
Nucl. Med.
52(6): 896-900, 2011; and William and Bierman Expert Op/n. Biol. Ther. 10(8):
1271-1278, 2010). Additional examples of CD20 antibodies are known in the art (see, e.g., WO
2008/156713).
In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of a bispecific antibody (e.g., XmAb13676; REGN1979 (Bannerji et al., Blood 128: 621, 2016; and Smith et al., Sci. Rep. 5: 17943, 2015); PRO131921 (Casulo et al., Cl/n. Immnol. 154(1): 37-46, 2014; and Robak and RobakBioDrugs 25(1): 13-25, 2011); or Acellbia).
In some embodiments, the CD20 inhibitor comprises or consists of a trispecific antibody (e.g., FBTA05 (Bi20; Lymphomun) (Buhmann et al., I Transl. Med.
11:160, 2013;
and Schuster et al., Br. I Haematol. 169(1): 90-102, 2015)).
Additional examples of CD20 inhibitors that are antibodies or antigen-binding fragments are described in, e.g., U.S. Patent Application Publication Nos.
2017/0304441, 2017/0128587, 2017/0088625, 2017/0037139, 2017/0002084, 2016/0362472, 2016/0347852, 2016/0333106, 2016/0271249, 2016/0243226, 2016/0115238, 2016/0108126, 2016/0017050, 2016/0017047, 2016/0000912, 2016/0000911, 2015/0344585, 2015/0290317, 2015/0274834, 2015/0265703, 2015/0259428, 2015/0218280, 2015/0125446, 2015/0093376, 2015/0079073, 2015/0071911, 2015/0056186, 2015/0010540, 2014/0363424, 2014/0356352, 2014/0328843, 2014/0322200, 2014/0294807, 2014/0248262, 2014/0234298, 2014/0093454, 2014/0065134, 2014/0044705, 2014/0004104, 2014/0004037, 2013/0280243, 2013/0273041, 2013/0251706, 2013/0195846, 2013/0183290, 2013/0089540, 2013/0004480, 2012/0315268, 2012/0301459, 2012/0276085, 2012/0263713, 2012/0258102, 2012/0258101, 2012/0251534, 2012/0219549, 2012/0183545, 2012/0100133, 2012/0034185, 2011/0287006, 2011/0263825, 2011/0243931, 2011/0217298, 2011/0200598, 2011/0195022, 2011/0195021, 2011/0177067, 2011/0165159, 2011/0165152, 2011/0165151, 2011/0129412, 2011/0086025, 2011/0081681, 2011/0020322, 2010/0330089, 2010/0310581, 2010/0303808, 2010/0183601, 2010/0080769, 2009/0285795, 2009/0203886, 2009/0197330, 2009/0196879, 2009/0191195, 2009/0175854, 2009/0155253, 2009/0136516, 2009/0130089, 2009/0110688, 2009/0098118, 2009/0074760, 2009/0060913, 2009/0035322, 2008/0260641, 2008/0213273, 2008/0089885, 2008/0044421, 2008/0038261, 2007/0280882, 2007/0231324, 2007/0224189, 2007/0059306, 2007/0020259, 2007/0014785, 2007/0014720, 2006/0121032, 2005/0180972, 2005/0112060, 2005/0069545, 2005/0025764, 2004/0213784, 2004/0167319, 2004/0093621, 2003/0219433, 2003/0206903, 2003/0180292, 2003/0026804, 2002/0039557, 2002/0012665, and 2001/0018041, each herein incorporated by reference in their entirety (e.g., sections describing CD20 inhibitors).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (KD) of less than 1 x 10-5M (e.g., less than 0.5 x 10-5M, less than 1 x 10-6M, less than 0.5 x 10-6M, less than 1 x 10-7M, less than 0.5 x 10-7M, less than 1 x 10-8M, less than 0.5 x 10-8M, less than 1 x 10-9M, less than 0.5 x 10-9M, less than 1 x 10-10 M, less than 0.5 x 10-10 M, less than 1 x 10-11M, less than 0.5 x 10-11M, or less than 1 x 10-12M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a KID of about 1 x 10-12M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10-6 M, about 0.5 x 10-6M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8 M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-10 M, about 0.5 x 10-10 M, about 1 x 10-11M, or about 0.5 x 10-11M (inclusive); about 0.5 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10-6M, about 0.5 x 10-6M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9 M, about 1 x 10-10 M, about 0.5 x 10-1 M, or about 1 x 10-11M (inclusive); about 1 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10-6M, about 0.5 x 10-6M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-10 M, or about 0.5 x 10-10 M (inclusive); about 0.5 x 10-10 M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10-6M, about 0.5 x 10-6M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, or about 1 x 10-1 M
(inclusive);
about 1 x 10-10 M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 106 M, about 0.5 x 10 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9 M, or about 0.5 x 10-9M (inclusive); about 0.5 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, or about 1 x 10-9M (inclusive); about 1 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-8M, or about 0.5 x 10-8M (inclusive); about 0.5 x 10-8M to about 1 x 10-5 M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, or about 1 x 10-8M (inclusive); about 1 x 10-8M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 106 M, about 0.5 x 10-6M, about 1 x 10' M, or about 0.5 x 10' M
(inclusive);
about 0.5 x 10' M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10-6M, about 0.5 x 10' M, or about 1 x 10-7M (inclusive); about 1 x 10-7M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10-6M, or about 0.5 x 10-6M (inclusive); about 0.5 x 10-6M to about 1 x 10-5M, about 0.5 x 10-5M, or about 1 x 10-6M (inclusive); about 1 x 10-6M to about 1 x 10-5M or about 0.5 x 10-5M (inclusive); or about 0.5 x 10-5M to about 1 x 10-5M
(inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Kar of about lx 10-6 s-1 to about lx 10-3 s-1, about 0.5 x 10-3 s-1, about lx 10-4 s-',about 0.5 x 10-4 s-1, about 1 x 10-5 s-1, or about 0.5 x 10-5 s-1 (inclusive); about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, about 0.5 x 10-4 s-1, or about 1 x 10-5 s-1 (inclusive); about 1 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, or about 0.5 x 10-4 s-1 (inclusive); about 0.5 x 10-4 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, or about 1 x 10-4 s-1 (inclusive); about 1 x 10-4 s-1 to about 1 x 10-3 s-1, or about 0.5 x 10-3 s-1 (inclusive); or about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Kon of about 1 x 102 M's' to about 1 x 106 N4-1S-1, about 0.5 x 106 M's', about 1 x i05 M's', about 0.5 x 10 M's', about 1 x 104 N4-1S-1, about 0.5 x iO4 M's', about 1 x 103 M's', or about 0.5 x 103 Ws-1 (inclusive); about 0.5 x 103M-1s-1-to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x i05 M's', about 0.5 x 10 M's', about 1 x iO4 M's', about 0.5 x 104 M's', or about 1 x iO3 M's' (inclusive); about 1 x iO3 M's' to about 1 x 106 M-1s-1, about 0.5 x 106 N4-1 s about 1 x 10 M's', about 0.5 x 10 M's', about 1 x 104 M's', or about 0.5 x 104 N4-1s-1 (inclusive); about 0.5 x 104 M's' to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105 M's', about 0.5 x 105 M's', or about 1 x 104 M"ls"1 (inclusive);
about 1 x 104N4-1s --1 to about 1 x 106 M's', about 0.5 x 106 m-ls-1, about 1 x 105 M's', or about 0.5 x 105 M"ls"1 (inclusive); about 0.5 x 105 M"ls"1 to about 1 x 106 M"1-s"1-, about 0.5 x s, or about 1 x 105 M"ls"1 (inclusive); about 1 x 105 M"ls"1 to about 1 x 106 M's', or about 0.5 x 106 m-ls-1 (inclusive); or about 0.5 x 106 M's' to about 1 x 106 M"ls"1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
Additional examples of CD20 inhibitors that are antibodies or antigen-binding fragments are known in the art.
CD20 Inhibitors- Peptides and Fusion Proteins In some embodiments, the CD20 inhibitor is an immunotoxin (e.g., MT-3724 (Hamlin Blood 128: 4200, 2016).
In some embodiments, the CD20 inhibitor is a fusion protein (e.g., TRU-015 (Rubbert-Roth Curr. Op/n. Mol. Ther 12(1): 115-123, 2010). Additional examples of CD20 inhibitors that are fusion proteins are described in, e.g., U.S. Patent Application Publication Nos. 2012/0195895, 2012/0034185, 2009/0155253, 2007/0020259, and 2003/0219433, each of which are herein incorporated by reference in their entirety (e.g., sections describing CD20 inhibitors).
CD25 Inhibitors The term "CD25 inhibitors" refers to an agent which decreases the ability of (also called interleukin-2 receptor alpha chain) to bind to interleukin-2.
CD25 forms a complex with interleukin-2 receptor beta chain and interleukin-2 common gamma chain.
In some embodiments, the CD25 inhibitor is an antibody or an antigen-binding fragment thereof, or a fusion protein. Exemplary CD25 inhibitors are described herein.
Additional examples of CD25 inhibitors are known in the art.
An exemplary sequence of human CD25 is shown below.
.. Human CD25 Isoform 1 (SEQ ID NO: 70) elcdddppe iphatfkama ykegtmlnce ckrgfrriks gslymlctgn sshsswdnqc qctssatmt tkqvtpqpee qkerkttemq spmqpvdqas 1pghcreppp weneateriy hfvvgqmvyy qcvqgyralh rgpaesvckm thgktrwtqp qlictgemet sqfpgeekpq aspegrpese tsclvtadf qiqtemaatm etsiftteyq vavagcvfll isylllsglt wqrrqrksrr ti Human CD25 Isoform 2 (SEQ ID NO: 71) elcdddppe iphatfkama ykegtmlnce ckrgfrriks gslymlctgn sshsswdnqc qctssatmt tkqvtpqpee qkerkttemq spmqpvdqas 1pgeekpqas pegrpesets clvtudfqi qtemaatmet siftteyqva vagcvfllis vlllsgltwq rrqrksrrti Human CD25 Isoform 3 (SEQ ID NO: 72) elcdddppe iphatfkama ykegtmlnce ckrgfrriks gslymlctgn sshsswdnqc qctssatmt tkqvtpqpee qkerkttemq spmqpvdqas 1pdfqiqtem aatmetsift teyqvavagc vfllisylllsgltwqrrqr ksrrti CD25 Inhibitors- Antibodies In some embodiments, the CD25 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv). In some embodiments, a CD25 inhibitor is an antibody or an antigen-binding fragment thereof that specifically binds to CD25. In some embodiments, a CD25 inhibitor is an antibody that specifically binds to IL-2.
In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv)2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat.
Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC
(Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO
Mol. Med.
9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFy (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFy (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, la-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG

(L,H)-Fc, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Patent No. 5,759,808; Stijlemans et al., I Biol.
Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., 1 Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol.
25(2):85-91, 2004), Diabody-CH3 (Guo et al.õ Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab')2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther.
8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.
Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab')2 fragment, and a Fab' fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgGl, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgGl, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgAl or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgAl or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).
In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986;
WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., I Exp. Med.
175:217-225, 1992; Kolstelny etal., I Immunol. 148(5):1547-1553, 1992; Hollinger etal., Proc. Natl.
Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., I Immunol. 152:5368, 1994; Tutt et al., Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC
Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (Fab' scFv)2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Patent No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., I Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.
In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Patent Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895;
US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US
2017/0002082, each of which is incorporated by reference in its entirety.
In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of basiliximab (SimulectTM) (Wang et al., Cl/n. Exp.
Immunol. 155(3):
496-503, 2009; and Kircher et al., Cl/n. Exp. Immunol. 134(3): 426-430, 2003);
daclizumab (Zenapax; Zinbrytag) (Berkowitz et al., Cl/n. Immunol. 155(2): 176-187, 2014;
and Bielekova et al., Arch Neurol. 66(4): 483-489, 2009); or IMTOX-25.
In some embodiments, the CD25 inhibitor is an antibody-drug-conjugate (e.g., ADCT-301 (Flynn et al., Blood 124: 4491, 2014)).
Additional examples of CD25 inhibitors that are antibodiesare known in the art (see, e.g., WO 2004/045512). Additional examples of CD25 inhibitors that are antibodies or antigen-binding fragments are described in, e.g., U.S. Patent Application Publication Nos.
2017/0240640, 2017/0233481, 2015/0259424, 2015/0010539, 2015/0010538, 2012/0244069, 2009/0081219, 2009/0041775, 2008/0286281, 2008/0171017, 2004/0170626, 2001/0041179, and 2010/0055098, each of which is incorporated herein by reference (e.g., sections that describe CD25 inhibitors).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (KD) of less than 1 x 10-5M (e.g., less than 0.5 x 10-5M, less than 1 x 10-6M, less than 0.5 x 10-6 M, less than 1 x 10-7M, less than 0.5 x 10-7M, less than 1 x 10-8M, less than 0.5 x 10-8M, less than 1 x 10-9M, less than 0.5 x 10-9M, less than 1 x 10-10 M, less than 0.5 x 10-10 M, less than 1 x 10-11M, less than 0.5 x 10-11M, or less than 1 x 1012M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a KID of about 1 x 1012 M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8 M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 1010 M, about 0.5 x 10-10 M, about 1 x 10-11M, or about 0.5 x 10-11M (inclusive); about 0.5 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9 M, about 1 x 10-10 M, about .. 0.5 x 10' M, or about 1 x 10-11M (inclusive); about 1 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 1010 M, or about 0.5 x 10-10 M (inclusive); about 0.5 x 10-10 M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, or about 1 x 1010 M
(inclusive);
about 1 x 1010 M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9 M, or about 0.5 x 10-9M (inclusive); about 0.5 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, or about 1 x 10-9M (inclusive); about 1 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, about 1 x 10-8M, or about 0.5 x 10-8M (inclusive); about 0.5 x 10-8M to about 1 x 10-5 M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, or about 1 x 10-8M (inclusive); about 1 x 10-8M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, or about 0.5 x 10-7M
(inclusive);
about 0.5 x 10-7M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, or about 1 x 10-7M (inclusive); about 1 x 10-7M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, or about 0.5 x 10' M (inclusive); about 0.5 x 10' M to about 1 x 10-5M, about 0.5 x 10-5M, or about 1 x 10' M (inclusive); about 1 x 10' M to about 1 x 10-5M or .. about 0.5 x 10-5M (inclusive); or about 0.5 x 10-5M to about 1 x 10-5M
(inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Koff of about lx 10' s-1 to about lx 10-3 s-1, about 0.5 x 10-3 s-1, about lx 10-4 s-1, about 0.5 x 10-4 s-1, about 1 x 10-5 s-1, or about 0.5 x 10-5 s-1 (inclusive); about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, about 0.5 x 10-4 s-1, or about 1 x 10-5 s-1 (inclusive); about 1 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, or about 0.5 x 10-4 s-1 (inclusive); about 0.5 x 10-4 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, or about 1 x 10-4 s-1 (inclusive); about 1 x 10-4 s-1 to about 1 x 10-3 s-1, or about 0.5 x 10-3 s-1 (inclusive); or about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Kon of about 1 x 102 M's' to about 1 x 106 N4-1S-1, about 0.5 x 106 M's', about 1 x i0 M's', about 0.5 x 10 M's', about 1 x 104 N4-1S-1, about 0.5 x iO4 M's', about 1 x 103 M's', or about 0.5 x iO3 M's' (inclusive); about 0.5 x iO3 M's' to about 1 x 106 N4-1s-1, about 0.5 x 106 M's', about 1 x i05 M's', about 0.5 x 10 M's', about 1 x iO4 M's', about 0.5 x 104 M's', or about 1 x 103M-1s-1 (inclusive); about 1 x 103M-1s-1 to about 1 x 106 M-1s-1, about 0.5 x 106 m-ls-1, about 1 x 10 M's', about 0.5 x 10 M's', about 1 x 104 M's', or about 0.5 x 104 N4-1s-1 (inclusive); about 0.5 x 104 ws-1 to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x i05 M's', about 0.5 x i05 M's', or about 1 x iO4 M's' (inclusive);
about 1 x 104N4-1s --1 to about 1 x 106 M's', about 0.5 x 106 Nrls-1, about 1 x 105 M's', or about 0.5 x 10 M's' (inclusive); about 0.5 x 10 M's' to about 1 x 106 M's', about 0.5 x s, or about 1 x i05 M's' (inclusive); about 1 x 10 M's' to about 1 x 106 M's', or about 0.5 x 106 m-ls-1 (inclusive); or about 0.5 x 106 ws-1 to about 1 x 106 M's' (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
Additional examples of CD25 inhibitors that are antibodies or antigen-binding fragments are known in the art.
CD25 Inhibitors- Fusion Proteins In some embodiments, the CD25 inhibitor is a fusion protein. See, e.g., Zhang et al., PNAS 100(4): 1891-1895, 2003.
CD28 Inhibitors The term "CD28 inhibitors" refers to an agent which decreases the ability of CD28 to bind to one or both of CD80 and CD86. CD28 is a receptor that binds to its ligands, CD80 (also called B7.1) and CD86 (called B7.2).

In some embodiments, the CD28 inhibitor can decrease the binding between CD28 and CD80 by blocking the ability of CD28 to interact with CD80. In some embodiments, the CD28 inhibitor can decrease the binding between CD28 and CD86 by blocking the ability of CD28 to interact with CD86. In some embodiments, the CD28 inhibitor can decrease the binding of CD28 to each of CD80 and CD86.
In some embodiments, the CD28 inhibitor is an antibody or an antigen-binding fragment thereof, a fusion protein, or peptide. Exemplary CD28 inhibitors are described herein. Additional examples of CD28 inhibitors are known in the art.
Exemplary sequences for human CD28, human CD80, and human CD86 are shown below.
Human CD28 Isoform 1 (SEQ ID NO: 73) nkilvkqspmlv aydnavnlsc kysynlfsre fraslhkgld savevcvvyg nysqqlqvys ktgfncdgkl gnesvtfylq nlyvnqtdiy fckievmypp pyldneksng tiihvkgkhl cpsplfpgps kpfwvlvvvg gvlacysllv tvafiifwvr skrsrllhsd ymnmtprrpg ptrkhyqpya pprdfaayrs Human CD28 Isoform 2 (SEQ ID NO: 74) nkilvkqspmlv aydnavnlsw khlcpsplfp gpskpfwvlv vvggvlacys llvtvafiif wvrskrsrll hsdymnmtpr rpgptrkhyq pyapprdfaa yrs Human CD28 Isoform 3 (SEQ ID NO: 75) khlcpsplfpgp skpfwvlvvv ggvlacysll vtvafiifwv rskrsrllhs dymnmtprrp gptrkhyqpy apprdfaayr s Human CD80 (SEQ ID NO: 76) vihvtk evkevatlsc ghnvsveela qtriywqkek kmvltmmsgd mniwpeyknr tifditnnls ivilalrpsd egtyecwlk yekdafkreh laevtlsvka dfptpsisdf eiptsnirri icstsggfpe phlswlenge elnainttvs qdpetelyav sskldfnmtt nhsfincliky ghlrvnqtfn wnttkqehfp dnllpswait lisvngifvi ccltycfapr crerrrnerl rresvrpv Human CD86 Isoform 1 (SEQ ID NO: 77) yfnetadlpc qfansqnqsl selvvfwqdq enlvinevyl gkekfdsvhs kymgrtsfds dswfirlhnl qikdkglyqc iihhkkptgm irihqmnsel svlanfsqpe ivpisniten vyinitcssi hgypepkkms vllrtknsti eydgimqksq dnvtelydvs islsysfpdv tsnmtifcil etdktrllss pfsieledpq pppdhipwit avlptvficv mvfclilwkw kkkkrprnsy kcgtntmere eseqtkkrek ihipersdea qrvfksskts scdksdtcf Human CD86 Isoform 2 (SEQ ID NO: 78) yfneta dlpcqfansq nqslselvvf wqdqenlvin evylgkekfd svhskymgrt sfdsdswtlr lhnlqikdkg lyqciihhkk ptgmirihqm nselsvlanf sqpeivpisn itenvyinit cssihgypep kkmsvllrtk nstieydgim qksqdnvtel ydvsislsys fpdvtsnmti fciletdktr llsspfsiel edpqpppdhi pwitavlptv iicvmvfcli lwkwkkkkrp rnsykcgtnt mereeseqtk krekihiper sdeaqrvfks sktsscdksd tcf Human CD86 Isoform 3 (SEQ ID NO: 79) yfneta dlpcqfansq nqslselvvf wqdqenlvin evylgkekfd svhskymgrt sfdsdswth lhnlqikdkg lyqciihhkk ptgmirihqm nselsvlanf sqpeivpisn itenvyinit cssihgypep kkmsvllrtk nstieydgim qksqdnvtel ydvsislsys fpdvtsnmti fciletdktr llsspfsigt ntmereeseq tkkrekihip ersdeaqrvf kssktsscdk sdtcf Human CD86 Isoform 4 (SEQ ID NO: 80) eiv pisnitenvy initcssihg ypepkkmsvl lrtknstiey dgimqksqdn vtelydvsis lsysfpdvts nmtifcilet dktrllsspf sieledpqpp pdhipwitav 1ptviicvmv fclilwkwkk kkrpmsykc gtntmerees eqtkkrekih ipersdeaqr vfkssktssc dksdtcf Human CD86 Isoform 5 (SEQ ID NO: 81) mgrtsfdsds wtlrlhnlqi kdkglyqcii hhkkptgmir ihqmnselsv lanfsqpeiv pisnitenvy initcssihg ypepkkmsvl lrtknstiey dgimqksqdn vtelydvsis lsysfpdvts nmtifcilet dktrllsspf sieledpqpp pdhipwitav 1ptviicvmv fclilwkwkk kkrpmsykc gtntmerees eqtkkrekih ipersdeaqr vfkssktssc dksdtcf CD28 Inhibitors- Antibodies In some embodiments, the CD28 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv).
In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunot 75:28-37, 2016), a (scFv)2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat.
Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC
(Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO
Mol. Med.
9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, la-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG
(L,H)-Fc, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Patent No. 5,759,808; Stijlemans et al., I Biol.
Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., 1 Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol.
25(2):85-91, 2004), Diabody-CH3 (Guo et al.õ Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab')2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther.
8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.
Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab')2 fragment, and a Fab' fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgGl, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgGl, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgAl or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgAl or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).
In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986;

WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby etal., I Exp. Med.
175:217-225, 1992; Kolstelny etal., I Immunol. 148(5):1547-1553, 1992; Hollinger etal., Proc. Natl.
Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., I Immunol. 152:5368, 1994; Tutt et al., Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC
Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (Fab' scFv)2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt etal., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Patent No. 4,676,980), a linear antibody (Zapata etal., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., I Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.
In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Patent Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895;
US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US
2017/0002082, each of which is incorporated by reference in its entirety.
In some embodiments, the CD28 inhibitor is a monovalent Fab' antibody (e.g., CFR104) (Poirier et al., Am. I Transplant 15(1): 88-100, 2015).
Additional examples of CD28 inhibitors that are antibodies or antigen-binding fragments are described in, e.g., U.S. Patent Application Publication Nos.
2017/0240636, 2017/0114136, 2016/0017039, 2015/0376278, 2015/0299321, 2015/0232558, 2015/0150968, 2015/0071916, 2013/0266577, 2013/0230540, 2013/0109846, 2013/0078257, 2013/0078236, 2013/0058933, 2012/0201814, 2011/0097339, 2011/0059071, 2011/0009602, 2010/0266605, 2010/0028354, 2009/0246204, 2009/0117135, 2009/0117108, 2008/0095774, 2008/0038273, 2007/0154468, 2007/0134240, 2007/0122410, 2006/0188493, 2006/0165690, 2006/0039909, 2006/0009382, 2006/0008457, 2004/0116675, 2004/0092718, 2003/0170232, 2003/0086932, 2002/0006403, 2013/0197202, 2007/0065436, 2003/0180290, 2017/0015747, 2012/0100139, and 2007/0148162, each of which is incorporated by reference in its entirety (e.g., sections that described CD28 inhibitors).

In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (KD) of less than 1 x 10-5M (e.g., less than 0.5 x 10-5M, less than 1 x 10' M, less than 0.5 x 10' M, less than 1 x 10-7M, less than 0.5 x 10-7M, less than 1 x 10-8M, less than 0.5 x 10-8M, less than 1 x 10-9M, less than 0.5 x 10-9M, less than 1 x 10-10 M, less than 0.5 x 10-10 M, less than 1 x 10-11M, less than 0.5 x 10-11M, or less than 1 x 10-12M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a KID of about 1 x 10-12M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8 M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-10 M, about 0.5 x 10-10 M, about 1 x 10-11M, or about 0.5 x 10-11M (inclusive); about 0.5 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9 M, about 1 x 10-10 M, about .. 0.5 x 10' M, or about 1 x 10-11M (inclusive); about 1 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-10 M, or about 0.5 x 10-10 M (inclusive); about 0.5 x 10-10 M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, or about 1 x 10-10 M
(inclusive);
about 1 x 10-10 M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9 M, or about 0.5 x 10-9M (inclusive); about 0.5 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, or about 1 x 10-9M (inclusive); about 1 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, about 1 x 10-8M, or about 0.5 x 10-8M (inclusive); about 0.5 x 10-8M to about 1 x 10-5 M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, or about 1 x 10-8M (inclusive); about 1 x 10-8M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, or about 0.5 x 10-7M
(inclusive);
about 0.5 x 10-7M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, or about 1 x 10-7M (inclusive); about 1 x 10-7M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, or about 0.5 x 10' M (inclusive); about 0.5 x 10' M to about 1 x 10-5M, about 0.5 x 10-5 M, or about 1 x 10' M (inclusive); about 1 x 10' M to about 1 x 10-5 M or about 0.5 x 10-5 M (inclusive); or about 0.5 x 10-5 M to about 1 x 10-5 M
(inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Kar of about lx 10' s-1 to about lx 10-3 s-1, about 0.5 x 10-3 s-1, about lx 10-4 s-1, about 0.5 x 10-4 s-1, about 1 x 10-5 s-1, or about 0.5 x 10-5 s-1 (inclusive); about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, about 0.5 x 10-4 s-1, or about 1 x 10-5 s-1 (inclusive); about 1 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, or about 0.5 x 10-4 s-1 (inclusive); about 0.5 x 10-4 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, or about 1 x 10-4 s-1 (inclusive); about 1 x 10-4 s-1 to about 1 x 10-3 s-1, or about 0.5 x 10-3 s-1 (inclusive); or about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Kon of about 1 x 102 M's' to about 1 x 106 M's', about 0.5 x 106 M's', about 1 .. x 105 M's', about 0.5 x 105 M's', about 1 x 104 N4-1S-1, about 0.5 x 104 M's', about 1 x 103 M's', or about 0.5 x 103 M-1s-1 (inclusive); about 0.5 x 103 M-1s-1 to about 1 x 106 N4-1s-1, about 0.5 x 106 M's', about 1 x 105 M's', about 0.5 x 105 M's', about 1 x 104 M's', about 0.5 x 104 M's', or about 1 x 103 M-1s-1 (inclusive); about 1 x 103 M-1s-1 to about 1 x 106 M-1s-1, about 0.5 x 106 m-ls-1, about 1 x 105 M's', about 0.5 x 105 M's', about 1 x 104 M's', or about 0.5 x 104 N4-1s-1 (inclusive); about 0.5 x 104 ws-1 to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105 M's', about 0.5 x 105 M's', or about 1 x 104 M-1s-1 (inclusive);
about 1 x 104 M's' to about 1 x 106 M's', about 0.5 x 106 Nrls-1, about 1 x 105 M's', or about 0.5 x 105 M-1s-1 (inclusive); about 0.5 x 105 M-1s-1 to about 1 x 106 M's', about 0.5 x 106 M's', or about 1 x 105 M-1s-1 (inclusive); about 1 x 105 M-1s-1 to about 1 x 106 M's', or about 0.5 x 106 m-ls-1 (inclusive); or about 0.5 x 106 ws-1 to about 1 x 106 M-1s-1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
Additional examples of CD28 inhibitors that are antibodies or antigen-binding fragments are known in the art.
CD28 Inhibitors- Fusion Proteins and Peptides In some embodiments, the CD28 inhibitor is a fusion protein (see, e.g., US
5,521,288;
and US 2002/0018783). In some embodiments, the CD28 inhibitor is abatacept (Orenciag) (Herrero-Beaumont et al., Rheumatol. Cl/n. 8: 78-83, 2012; and Korhonen and Moilanen Basic Clin. Pharmacol. Toxicol. 104(4): 276-284, 2009).
In some embodiments, the CD28 inhibitor is a peptide mimetic (e.g., AB103) (see, e.g., Bulger et al., AMA Surg. 149(6): 528-536, 2014), or a synthetical peptoid (see, e.g.., Li et al., Cell Mol. Immunol. 7(2): 133-142, 2010).
CD49 Inhibitors The term "CD49 inhibitors" refers to an agent which decreases the ability of CD49 to bind to one of its ligands (e.g., MMP1).. In some embodiments, the CD49 inhibitor is an antibody or an antigen-binding fragment thereof. Exemplary CD49 inhibitors are described herein. Additional examples of CD49 inhibitors are known in the art.
Exemplary sequences for human CD49 and human MMP1 are shown below.
Human CD49 (SEQ ID NO: 82) mgpertgaaplpillylals qgilncclay nvglpeakif sgpsseqfgy avqqfinpkg nwllvgspws gfpenrmgdv ykcpvdlsta tceklnlqts tsipnvtemk tnmslglilt rnmgtggflt cgplwaqqcg nqyyttgvcs dispdfqlsa sfspatqpcp slidvvvvcd esnsiypwda yknflekfyq gldigptktq vgliqyannp rvvfnlntyk tkeemivats qtsqyggdlt ntfgaiqyar kyaysaasgg rrsatkvmvy vtdgeshdgs mlkavidqcn hdnilrfgia vlgylnrnal dtknlikeik aiasiptery ffnvsdeaal lekagtlgeq ifsiegtvqg gdnfqmemsq vgfsadyssq ndilmlgavg afgwsgtivq ktshghlifp kqafdqilqd rnhssylgys vaaistgest hfvagapran ytgqivlysv nengnitviq ahrgdqigsy fgsylcsydy dkdfitdyll vgapmymsdl kkeegrvylf tikkgilgqh qflegpegie ntrfgsaiaa lsdinmdgfn dvivgsplen qnsgavyiyn ghqgtirtky sqkilgsdga frshlqyfgr sldgygdlng dsitdvsiga fgqvvqlwsq siadvaieas ftpekitivn knaqiilklc fsakfrptkq nnqvaivyni tldadgfssr vtsrglfken nerclqknmv vnqaqscpeh iiyiqepsdv vnsldlrydi slenpgtspa leaysetakv fsipfhkdcg edglcisdlv ldvrqipaaq eqpfivsnqn krltfsvtlk nkresayntg ivvdfsenlf fasfslpvdg tevtcqvaas qksvacdvgy palkreqqvt ftinfdfnlq nlqnqaslsf qalsesqeen kadnlvnlki pllydaeihl trstninfye issdgnvpsi vhsfedvgpk fifslkvttg svpvsmatvi ihipqytkek nplmyltgvq tdkagdiscn adinplkigq tsssysfkse nfrhtkelnc rtascsnvtc wlkdvhmkge yfvnyttriw ngtfasstfq tvqltaaaei ntynpeiyvi edntytiplm imkpdekaev ptgviigsii agillllalv ailwklgffk rkyekmtknp deidettels s Human MMP1 (SEQ ID NO: 83) mhsfpp11111fwgyvshsf patletqeqd vdlyqkylek yynlkndgrq vekrrnsgpv veklkqmqef fglkvtgkpd aetlkvmkqp rcgvpdvaqf vltegnprwe qthltyrien ytpdlpradv dhaiekafql wsnvtpltft kvsegqadim isfvrgdhrd nspfdgpggn lahafqpgpg iggdahfded erwtnnfrey nlhrvaahel ghslglshst digalmypsy tfsgdyglaq ddidgiqaiy grsqnpvqpi gpqtpkacds kltfdaitti rgevmffkdr fymrtnpfyp evelnfisvf wpqlpnglea aye fadrdev rffkgnkywa vqgqnvlhgy pkdiyssfgf prtvkhidaa lseentgkty ffvankywry deykrsmdpg ypkmiandfp gighkvdavf mkdgffyffh gtrqykfdpk tkriltlqka nswfncrkn CD49 Inhibitors- Antibodies In some embodiments, the CD49 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv).
In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunol. 75:28-37, 2016), a (scFv)2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat.
Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC
(Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula et al., EMBO
Mol. Med.
9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume et al., Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, la-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG
(L,H)-Fc, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Patent No. 5,759,808; Stijlemans et al., I Biol.
Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al., 1 Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol.
25(2):85-91, 2004), Diabody-CH3 (Guo et al.õ Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab')2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston et al., Human Antibodies 10(3-4):127-142, 2001; Wheeler et al., Mol. Ther.
8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.
Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab')2 fragment, and a Fab' fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgGl, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgGl, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgAl or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgAl or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).
In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986;
WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby et al., I Exp. Med.
175:217-225, 1992; Kolstelny et al., I Immunol. 148(5):1547-1553, 1992; Hollinger et al., Proc. Natl.
Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., I Immunol. 152:5368, 1994; Tutt et al., 1 Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC
Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (Fab' scFv)2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt et al., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Patent No. 4,676,980), a linear antibody (Zapata et al., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., I Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.

In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments thereof are described in U.S. Patent Nos. 8,440,196; 7,842,144; 8,034,344; and 8,529,895;
US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US
2017/0002082, each of which is incorporated by reference in its entirety.
In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of natalizumab (Tysabrig; Antegreng) (see, e.g., Pagnini et al., Expert Op/n. Biol. Ther. 17(11): 1433-1438, 2017; and Chataway and Miller Neurotherapeutics 10(1): 19-28, 2013; or vatelizumab (ELND-004)).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (KD) of less than 1 x 10-5M (e.g., less than 0.5 x 10-5M, less than 1 x 10' M, less than 0.5 x 10' M, less than 1 x 10-7M, less than 0.5 x 10-7M, less than 1 x 10-8M, less than 0.5 x 10-8M, less than 1 x 10-9M, less than 0.5 x 10-9M, less than 1 x 10-10 M, less than 0.5 x 10-10 M, less than 1 x 10-11M, less than 0.5 x 10-11M, or less than 1 x 10-12M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a KID of about 1 x 10-12M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8 M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-10 M, about 0.5 x 10-10 M, about 1 x 10-11M, or about 0.5 x 10-11M (inclusive); about 0.5 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7 M, about 0.5 x 10-7 M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9 M, about 1 x 10-10 M, about 0.5 x 10-1 M, or about 1 x 10-11M (inclusive); about 1 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 10-10 M, or about 0.5 x 10-10 M (inclusive); about 0.5 x 10-10 M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7 M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, or about 1 x 10-10 M
(inclusive);
about 1 x 10-10 M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9 M, or about 0.5 x 10-9M (inclusive); about 0.5 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-8 M, about 0.5 x 10-8M, or about 1 x 10-9M (inclusive); about 1 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, about 1 x 10-8M, or about 0.5 x 10-8M (inclusive); about 0.5 x 10-8M to about 1 x 10-5 M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10' M, about 0.5 x 10-M, or about 1 x 10-8M (inclusive); about 1 x 10-8M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, or about 0.5 x 10-7M
(inclusive);
.. about 0.5 x 10' M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, or about 1 x 10' M (inclusive); about 1 x 10' M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, or about 0.5 x 10' M (inclusive); about 0.5 x 10' M to about 1 x 10-5M, about 0.5 x 10-5M, or about 1 x 10' M (inclusive); about 1 x 10' M to about 1 x 10-5M or about 0.5 x 10-5M (inclusive); or about 0.5 x 10-5M to about 1 x 10-5M
(inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Koff of about lx 10' s-1 to about lx 10-3 s-1, about 0.5 x 10-3 s-1, about lx 10-4 s-1, about 0.5 x 10-4 s-1, about 1 x 10-5 s-1, or about 0.5 x 10-5 s-1 (inclusive); about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, about 0.5 x 10-4 s-1, or about 1 x 10-5 s-1 (inclusive); about 1 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s"
',or about 0.5 x 10-4 s-1 (inclusive); about 0.5 x 10-4 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s"
1, or about 1 x 10-4 s-1 (inclusive); about 1 x 10-4 s-1 to about 1 x 10-3 s-1, or about 0.5 x 10-3 s-1 (inclusive); or about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Kon of about 1 x 102 M's' to about 1 x 106 N4-1S-1, about 0.5 x 106 M's', about 1 x i0 M's', about 0.5 x 10 M's', about 1 x 104 N4-1S-1, about 0.5 x iO4 M's', about 1 x 103 M's', or about 0.5 x iO3 M's' (inclusive); about 0.5 x 103M-1s-1-to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x i05 M's', about 0.5 x 10 M's', about 1 x iO4 M's', about 0.5 x 104 M's', or about 1 x 103M-1s-1 (inclusive); about 1 x 103M-1s-1 to about 1 x 106 M-1s-1, about 0.5 x 106 m-ls-1, about 1 x 10 M's', about 0.5 x 10 M's', about 1 x 104M-1s-1, or about 0.5 x 104 N4-1s-1 (inclusive); about 0.5 x 104 ws-1 to about 1 X 106 M's', about 0.5 x 106 M's', about 1 x 105M-1s-1, about 0.5 x 105M-1s-1, or about 1 x 104M-1s-1 (inclusive);

about 1 x 104 M's' to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105 M's', or about 0.5 x i05 M's' (inclusive); about 0.5 x 105 M's' to about 1 x 106 M's', about 0.5 x 106 M's', or about 1 x 105 M's' (inclusive); about 1 x i05 M's' to about 1 x 106 M's', or about 0.5 x 106 M's' (inclusive); or about 0.5 x 106 M's' to about 1 x 106 M's' (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
Additional examples of CD49 inhibitors that are antibodies or antigen-binding fragments are known in the art.
CD89 Inhibitors The term "CD89 inhibitors" refers to an agent which decreases the ability of CD89 to bind to IgA. CD89 is a transmembrane glycoprotein that binds to the heavy-chain constant region of IgA. In some embodiments, the CD89 inhibitor can decrease the binding between CD89 and IgA by blocking the ability of CD89 to interact with IgA. In some embodiments, the CD89 inhibitor is an antibody or an antigen-binding fragment thereof.
Exemplary CD89 inhibitors are described herein. Additional examples of CD89 inhibitors are known in the art.
An exemplary sequence for human CD89 is shown below.
Human CD89 (SEQ ID NO: 84) mdpkqttilc lviclgqriq aqegdfpmpf isaksspvip ldgsvkiqcq aireayltql miiknstyre igrrlkfwne tdpefvidhm dankagryqc qyrighyrfr ysdtlelvvt glygkpflsa drglvlmpge nisltcssah ipfdrfslak egelslpqhq sgehpanfsl gpvdlnvsgi yrcygwynrs pylwsfpsna lelvvtdsih qdyttqnlir mavaglvlva llailvenwh shtalnkeas advaepswsq qmcqpgltfa rtpsvck CD89 Inhibitors- Antibodies In some embodiments, the CD89 inhibitor is an antibody or an antigen-binding fragment thereof (e.g., a Fab or a scFv).
In some embodiments, the antibody can be a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof. In some embodiments, an antibody can be a scFv-Fc (Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017), a VHH domain (Li et al., Immunol. Lett. 188:89-95, 2017), a VNAR domain (Hasler et al., Mol. Immunot 75:28-37, 2016), a (scFv)2, a minibody (Kim et al., PLoS One 10(1):e113442, 2014), or a BiTE. In some embodiments, an antibody can be a DVD-Ig (Wu et al., Nat.
Biotechnol. 25(11):1290-1297, 2007; WO 08/024188; WO 07/024715), and a dual-affinity re-targeting antibody (DART) (Tsai et al., Mol. Ther. Oncolytics 3:15024, 2016), a triomab (Chelius et al., MAbs 2(3):309-319, 2010), kih IgG with a common LC
(Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a crossmab (Regula etal., EMBO Mol.
Med.
9(7):985, 2017), an ortho-Fab IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a 2-in-1-IgG (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), IgG-scFv (Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014), scFv2-Fc (Natsume etal., Biochem. 140(3):359-368, 2006), a bi-nanobody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), tanden antibody (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a DART-Fc (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), a scFv-HSA-scFv (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DNL-Fab3 (Kontermann et al., Drug Discovery Today 20(7):838-847, 2015), DAF (two-in-one or four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair antibody, Fab-arm exchange antibody, SEEDbody, Triomab, LUZ-Y, Fcab, la-body, orthogonal Fab, DVD-IgG, IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)-IgG, IgG
(L,H)-Fc, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, Zybody, DVI-IgG, nanobody (e.g., antibodies derived from Camelus bactriamus, Calelus dromaderius, or Lama paccos) (U.S. Patent No. 5,759,808; Stijlemans et al., I Biol.
Chem. 279:1256-1261, 2004; Dumoulin et al., Nature 424:783-788, 2003; and Pleschberger et al., Bioconjugate Chem. 14:440-448, 2003), nanobody-HSA, a diabody (e.g., Poljak, Structure 2(12):1121-1123, 1994; Hudson et al.,1 Immunol. Methods 23(1-2):177-189, 1999), a TandAb (Reusch et al., mAbs 6(3):727-738, 2014), scDiabody (Cuesta et al., Trends in Biotechnol. 28(7):355-362, 2010), scDiabody-CH3 (Sanz et al., Trends in Immunol.
25(2):85-91, 2004), Diabody-CH3 (Guo et al.õ Triple Body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab')2-scFV2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intrabody (Huston etal., Human Antibodies 10(3-4):127-142, 2001; Wheeler etal., Mol. Ther.
8(3):355-366, 2003; Stocks, Drug Discov. Today 9(22):960-966, 2004), dock and lock bispecific antibody, ImmTAC, HSAbody, scDiabody-HSA, tandem scFv, IgG-IgG, Cov-X-Body, and scFv1-PEG-scFv2.
Non-limiting examples of an antigen-binding fragment of an antibody include an Fv fragment, a Fab fragment, a F(ab')2 fragment, and a Fab' fragment. Additional examples of an antigen-binding fragment of an antibody is an antigen-binding fragment of an IgG (e.g., an antigen-binding fragment of IgGl, IgG2, IgG3, or IgG4) (e.g., an antigen-binding fragment of a human or humanized IgG, e.g., human or humanized IgGl, IgG2, IgG3, or IgG4); an antigen-binding fragment of an IgA (e.g., an antigen-binding fragment of IgAl or IgA2) (e.g., an antigen-binding fragment of a human or humanized IgA, e.g., a human or humanized IgAl or IgA2); an antigen-binding fragment of an IgD (e.g., an antigen-binding fragment of a human or humanized IgD); an antigen-binding fragment of an IgE (e.g., an antigen-binding fragment of a human or humanized IgE); or an antigen-binding fragment of an IgM (e.g., an antigen-binding fragment of a human or humanized IgM).
In some embodiments, an antibody can be an IgNAR, a bispecific antibody (Milstein and Cuello, Nature 305:537-539, 1983; Suresh et al., Methods in Enzymology 121:210, 1986;
WO 96/27011; Brennan et al., Science 229:81, 1985; Shalaby etal., I Exp. Med.
175:217-.. 225, 1992; Kolstelny etal., I Immunol. 148(5):1547-1553, 1992; Hollinger etal., Proc. Natl.
Acad. Sci. U.S.A. 90:6444-6448, 1993; Gruber et al., I Immunol. 152:5368, 1994; Tutt et al., Immunol. 147:60, 1991), a bispecific diabody, a triabody (Schoonooghe et al., BMC
Biotechnol. 9:70, 2009), a tetrabody, scFv-Fc knobs-into-holes, a scFv-Fc-scFv, a (Fab' scFv)2, a V-IgG, a IvG-V, a dual V domain IgG, a heavy chain immunoglobulin or a camelid (Holt etal., Trends Biotechnol. 21(11):484-490, 2003), an intrabody, a monoclonal antibody (e.g., a human or humanized monoclonal antibody), a heteroconjugate antibody (e.g., U.S. Patent No. 4,676,980), a linear antibody (Zapata etal., Protein Eng. 8(10:1057-1062, 1995), a trispecific antibody (Tutt et al., I Immunol. 147:60, 1991), a Fabs-in-Tandem immunoglobulin (WO 15/103072), or a humanized camelid antibody.
In some embodiments, the antibody is a humanized antibody, a chimeric antibody, a multivalent antibody, or a fragment thereof In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized monoclonal antibody. See e.g., Hunter & Jones, Nat. Immunol. 16:448-457, 2015; Heo et al., Oncotarget 7(13):15460-15473, 2016. Additional examples of antibodies and antigen-binding fragments .. thereof are described in U.S. Patent Nos. 8,440,196; 7,842,144; 8,034,344;
and 8,529,895;
US 2013/0317203; US 2014/0322239; US 2015/0166666; US 2016/0152714; and US
2017/0002082, each of which is incorporated by reference in its entirety.
In certain embodiments, the antibody comprises or consists of an antigen-binding fragment or portion of HF-1020. Additional examples of CD89 antibodies are known in the art (see, e.g., WO 2002/064634).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a dissociation constant (KD) of less than 1 x 10-5M (e.g., less than 0.5 x 10-5M, less than 1 x 10' M, less than 0.5 x 10' M, less than 1 x 10-7M, less than 0.5 x 10-7M, less than 1 x 10-8M, less than 0.5 x 10-8M, less than 1 x 10-9M, less than 0.5 x 10-9M, less than 1 x 10-10 M, less than 0.5 x 10-10 M, less than 1 x 10-11M, less than 0.5 x 10-11M, or less than 1 x 10-12M), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a KID of about 1 x 10-12M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8 M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 1010 M, about 0.5 x 10-10 M, about 1 x 10-11M, or about 0.5 x 10-11M (inclusive); about 0.5 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9 M, about 1 x 1010 M, about 0.5 x 10' M, or about 1 x 10-11M (inclusive); about 1 x 10-11M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, about 1 x 1010 M, or about 0.5 x 10-10 M (inclusive); about 0.5 x 10-10 M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, about 1 x 10-9M, about 0.5 x 10-9M, or about 1 x 10-10 M
(inclusive);
about 1 x 1010 M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8M, about 0.5 x 10-8M, about 1 x 10-9 M, or about 0.5 x 10-9M (inclusive); about 0.5 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5 M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7M, about 1 x 10-8 M, about 0.5 x 10-8M, or about 1 x 10-9M (inclusive); about 1 x 10-9M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, about 1 x 10-8M, or about 0.5 x 10-8M (inclusive); about 0.5 x 10-8M to about 1 x 10-5 M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, about 0.5 x 10-7 M, or about 1 x 10-8M (inclusive); about 1 x 10-8M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, about 1 x 10-7M, or about 0.5 x 10-7M
(inclusive);
about 0.5 x 10-7M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, about 0.5 x 10' M, or about 1 x 10-7M (inclusive); about 1 x 10-7M to about 1 x 10-5M, about 0.5 x 10-5M, about 1 x 10' M, or about 0.5 x 10' M (inclusive); about 0.5 x 10' M to about 1 x 10-5M, about 0.5 x 10-5M, or about 1 x 10' M (inclusive); about 1 x 10' M to about 1 x 10-5M or about 0.5 x 10-5M (inclusive); or about 0.5 x 10-5M to about 1 x 10-5M
(inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).

In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Kar of about lx 10-6 s-1 to about lx 10-3 s-1, about 0.5 x 10-3 s-1, about lx 10-4 s-1, about 0.5 x 10-4 s-1, about 1 x 10-5 s-1, or about 0.5 x 10-5 s-1 (inclusive); about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, about 0.5 x 10-4 s-1, or about 1 x 10-5 s-1 (inclusive); about 1 x 10-5 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, about 1 x 10-4 s-1, or about 0.5 x 10-4 s-1 (inclusive); about 0.5 x 10-4 s-1 to about 1 x 10-3 s-1, about 0.5 x 10-3 s-1, or about 1 x 10-4 s-1 (inclusive); about 1 x 10-4 s-1 to about 1 x 10-3 s-1, or about 0.5 x 10-3 s-1 (inclusive); or about 0.5 x 10-5 s-1 to about 1 x 10-3 s-1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
In some embodiments, any of the antibodies or antigen-binding fragments described herein has a Kon of about 1 x 102 M's' to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105 M's', about 0.5 x 105 M's', about 1 x 104 N4-1S-1, about 0.5 x 104 M's', about 1 x 103 M's', or about 0.5 x 103 M-1s-1 (inclusive); about 0.5 x 103 M-1s-1 to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105 M's', about 0.5 x 105 M's', about 1 x 104 M's', about 0.5 x 104 M's', or about 1 x 103 M-1s-1 (inclusive); about 1 x 103 M-1s-1 to about 1 x 106 M-1s-1, about 0.5 x 106 m-ls-1, about 1 x 105 M's', about 0.5 x 105 M's', about 1 x 104 M's', or or about 0.5 x 104 (inclusive); about 0.5 x 104 ws-1 to about 1 x 106 M's', about 0.5 x 106 M's', about 1 x 105 M's', about 0.5 x 105 M's', or about 1 x 104 M-1s-1 (inclusive);
about 1 x 104 M's' to about 1 x 106 M's', about 0.5 x 106 m-ls-1, about 1 x 105 M's', or about 0.5 x 105 M-1s-1 (inclusive); about 0.5 x 105 M-1s-1 to about 1 x 106 M's', about 0.5 x 106 M's', or about 1 x 105 M-1s-1 (inclusive); about 1 x 105 M-1s-1 to about 1 x 106 M's', or about 0.5 x 106 m-ls-1 (inclusive); or about 0.5 x 106 ws-1 to about 1 x 106 M-1s-1 (inclusive), e.g., as measured in phosphate buffered saline using surface plasmon resonance (SPR).
Additional examples of CD89 inhibitors that are antibodies or antigen-binding fragments are known in the art.
CD283 (TLR3) Antibodies In some embodiments, the therapeutic agent is PRV-300, for example, as described in PCT publication WO 2006/060513 which is incorporated by reference herein in its entirety.
PRV-300 is an anti-Toll-Like Receptor 3 (TLR3)/CD283 monoclonal antibody that blocks TLR3 on cell surfaces and in endosomes.

IL-1 Inhibitors The term "IL-1 inhibitor" refers to an agent that decreases the expression of an IL-1 cytokine or an IL-1 receptor and/or decreases the ability of an IL-1 cytokine to bind specifically to an IL-1 receptor. Non-limiting examples of IL-1 cytokines include IL-la, IL-1(3, IL-18, IL-36a, IL-3613, IL-36y, IL-38, and IL-33. In some examples, an IL-1 cytokine is IL-la. In some examples, an IL-1 cytokine is IL-113.
As is known in the art, IL-la and IL-113 each binds to a complex of IL-1R1 and IL1RAP proteins; IL-18 binds to IL-18Ra; IL-36a, IL-3613, and IL-36y each binds to a complex of IL-1RL2 and IL-1RAP proteins; and IL-33 binds to a complex of IL1RL1 and IL1RAP proteins. IL-1Ra is an endogenous soluble protein that decreases the ability of IL-la and IL-113 to bind to their receptor (e.g., a complex of IL-1R1 and IL1RAP
proteins). IL-36Ra is an endogenous soluble protein that decreases the ability of IL-36a, IL-3613, and IL-36y to bind to their receptor (e.g., a complex of IL-1RL2 and IL-1RAP
proteins).
In some embodiments, the IL-1 inhibitor mimicks native human interleukin 1 receptor antagonist (ILl-Ra).
In some embodiments, the IL-1 inhibitor targets IL-la. In some embodiments, the IL-1 inhibitor targets IL-1(3. In some embodiments, the IL-1 inhibitor targets one or both of IL-1R1 and IL1RAP. For example, an IL-1 inhibitor can decrease the expression of IL-la and/or decrease the ability of IL-la to bind to its receptor (e.g., a complex of IL-1R1 and IL1RAP
proteins). In another example, an IL-1 inhibitor can decrease the expression of IL-113 and/or decrease the ability of IL-113 to binds to its receptor (e.g., a complex of IL-1R1 and IL1RAP
proteins). In some embodiments, an IL-1 inhibitor can decrease the expression of one or both of IL-1R1 and IL1RAP.
In some embodiments, the IL-1 inhibitor targets IL-18. In some embodiments, the IL-1 inhibitor targets IL-18Ra. In some embodiments, the IL-1 inhibitor decreases the ability of IL-18 to bind to its receptor (e.g., IL-18Ra). In some embodiments, the IL-1 inhibitor decreases the expression of IL-18. In some embodiments, the IL-1 inhibitor decreases the expression of IL-18Ra.
In some embodiments, the IL-1 inhibitor targets one or more (e.g., two or three) of IL-36a, IL-3613, and IL-36y. In some embodiments, the IL-1 inhibitor targets one or both of IL-1RL2 and IL-1RAP. In some embodiments, the IL-1 inhibitor decreases the expression of one or more (e.g., two or three) of IL-36a, IL-36(3, and IL-36y. In some embodiments, the IL-1 inhibitor decreases the expression of one or both of IL-1RL2 and IL-1RAP
proteins. In some embodiments, the IL-1 inhibitor decreases the ability of IL-36a to bind to its receptor (e.g., a complex including IL-1RL2 and IL-1RAP). In some examples, the IL-1 inhibitor decreases the ability of IL-3613 to bind to its receptor (e.g., a complex including IL-1RL2 and IL-1RAP). In some examples, the IL-1 inhibitor decreases the ability of IL-367 to bind to its receptor (e.g., a complex including IL-1RL2 and IL-1RAP).
In some embodiments, the IL-1 inhibitor targets IL-33. In some embodiments, the IL-1 inhibitor targets one or both of IL1RL1 and IL1RAP. In some embodiments, the inhibitor decreases the expression of IL-33. In some embodiments, the IL-1 inhibitor decreases the expression of one or both of IL1RL1 and IL1RAP. In some embodiments, the IL-1 inhibitor decreases the ability of IL-33 to bind to its receptor (e.g., a complex of IL1RL1 and IL1RAP proteins).
In some embodiments, an IL-1 inhibitory agent is an inhibitory nucleic acid, an antibody or fragment thereof, or a fusion protein. In some embodiments, the inhibitory nucleic acid is an antisense nucleic acid, a ribozyme, or a small interfering RNA.
Inhibitory Nucleic Acids Inhibitory nucleic acids that can decrease the expression of IL-la, IL-1(3, IL-18, IL-36a, IL-3613, IL-36y, IL-38, IL-33, IL-1R1, IL1RAP, IL-18Ra, IL-1RL2, or IL1RL1 mRNA
expression in a mammalian cell include antisense nucleic acid molecules, i.e., nucleic acid molecules whose nucleotide sequence is complementary to all or part of an IL-la, IL-113, IL-18, IL-36a, IL-3613, IL-36y, IL-38, IL-33, IL-1R1, IL1RAP, IL-18Ra, IL-1RL2, or IL1RL1 mRNA (e.g., complementary to all or a part of any one of SEQ ID NOs: 85-125).
Human IL-la mRNA (SEQ ID NO: 85) 1 agtaaccagg caacaccatt gaaggctcat atgtaaaaat ccatgccttc ctttctccca 61 atctccattc ccaaacttag ccactggctt ctggctgagg ccttacgcat acctcccggg 121 gcttgcacac accttcttct acagaagaca caccttgggc atatcctaca gaagaccagg 181 cttctctctg gtccttggta gagggctact ttactgtaac agggccaggg tggagagttc 241 tctcctgaag ctccatcccc tctataggaa atgtgttgac aatattcaga agagtaagag 301 gatcaagact tctttgtgct caaataccac tgttctcttc tctaccctgc cctaaccagg 361 agcttgtcac cccaaactct gaggtgattt atgccttaat caagcaaact tccctcttca 421 gaaaagatgg ctcattttcc ctcaaaagtt gccaggagct gccaagtatt ctgccaattc 481 accctggagc acaatcaaca aattcagcca gaacacaact acagctacta ttagaactat 541 tattattaat aaattcctct ccaaatctag ccccttgact tcggatttca cgatttctcc 601 cttcctccta gaaacttgat aagtttcccg cgcttccctt tttctaagac tacatgtttg 661 tcatcttata aagcaaaggg gtgaataaat gaaccaaatc aataacttct ggaatatctg 721 caaacaacaa taatatcagc tatgccatct ttcactattt tagccagtat cgagttgaat 781 gaacatagaa aaatacaaaa ctgaattctt ccctgtaaat tccccgtttt gacgacgcac 841 ttgtagccac gtagccacgc ctacttaaga caattacaaa aggcgaagaa gactgactca 901 ggcttaagct gccagccaga gagggagtca tttcattggc gtttgagtca gcaaagaagt 961 caagatggcc aaagttccag acatgtttga agacctgaag aactgttaca gtgaaaatga 1021 agaagacagt tcctccattg atcatctgtc tctgaatcag aaatccttct atcatgtaag 1081 ctatggccca ctccatgaag gctgcatgga tcaatctgtg tctctgagta tctctgaaac 1141 ctctaaaaca tccaagctta ccttcaagga gagcatggtg gtagtagcaa ccaacgggaa 1201 ggttctgaag aagagacggt tgagtttaag ccaatccatc actgatgatg acctggaggc 1261 catcgccaat gactcagagg aagaaatcat caagcctagg tcagcacctt ttagcttcct 1321 gagcaatgtg aaatacaact ttatgaggat catcaaatac gaattcatcc tgaatgacgc 1381 cctcaatcaa agtataattc gagccaatga tcagtacctc acggctgctg cattacataa 1441 tctggatgaa gcagtgaaat ttgacatggg tgcttataag tcatcaaagg atgatgctaa 1501 aattaccgtg attctaagaa tctcaaaaac tcaattgtat gtgactgccc aagatgaaga 1561 ccaaccagtg ctgctgaagg agatgcctga gatacccaaa accatcacag gtagtgagac 1621 caacctcctc ttcttctggg aaactcacgg cactaagaac tatttcacat cagttgccca 1681 tccaaacttg tttattgcca caaagcaaga ctactgggtg tgcttggcag gggggccacc 1741 ctctatcact gactttcaga tactggaaaa ccaggcgtag gtctggagtc tcacttgtct 1801 cacttgtgca gtgttgacag ttcatatgta ccatgtacat gaagaagcta aatcctttac 1861 tgttagtcat ttgctgagca tgtactgagc cttgtaattc taaatgaatg tttacactct 1921 ttgtaagagt ggaaccaaca ctaacatata atgttgttat ttaaagaaca ccctatattt 1981 tgcatagtac caatcatttt aattattatt cttcataaca attttaggag gaccagagct 2041 actgactatg gctaccaaaa agactctacc catattacag atgggcaaat taaggcataa 2101 gaaaactaag aaatatgcac aatagcagtt gaaacaagaa gccacagacc taggatttca 2161 tgatttcatt tcaactgttt gccttctact tttaagttgc tgatgaactc ttaatcaaat 2221 agcataagtt tctgggacct cagttttatc attttcaaaa tggagggaat aatacctaag 2281 ccttcctgcc gcaacagttt tttatgctaa tcagggaggt cattttggta aaatacttct 2341 tgaagccgag cctcaagatg aaggcaaagc acgaaatgtt attttttaat tattatttat 2401 atatgtattt ataaatatat ttaagataat tataatatac tatatttatg ggaacccctt 2461 catcctctga gtgtgaccag gcatcctcca caatagcaga cagtgttttc tgggataagt 2521 aagtttgatt tcattaatac agggcatttt ggtccaagtt gtgcttatcc catagccagg 2581 aaactctgca ttctagtact tgggagacct gtaatcatat aataaatgta cattaattac 2641 cttgagccag taattggtcc gatctttgac tcttttgcca ttaaacttac ctgggcattc 2701 ttgtttcaat tccacctgca atcaagtcct acaagctaaa attagatgaa ctcaactttg 2761 acaaccatga gaccactgtt atcaaaactt tcttttctgg aatgtaatca atgtttcttc 2821 taggttctaa aaattgtgat cagaccataa tgttacatta ttatcaacaa tagtgattga 2881 tagagtgtta tcagtcataa ctaaataaag cttgcaacaa aattctctga caaaaaaaaa 2941 aaaaaaa Human IL-1I3 mRNA (SEQ ID NO: 86) 1 accaaacctc ttcgaggcac aaggcacaac aggctgctct gggattctct tcagccaatc 61 ttcattgctc aagtgtctga agcagccatg gcagaagtac ctgagctcgc cagtgaaatg 121 atggcttatt acagtggcaa tgaggatgac ttgttctttg aagctgatgg ccctaaacag 181 atgaagtgct ccttccagga cctggacctc tgccctctgg atggcggcat ccagctacga 241 atctccgacc accactacag caagggcttc aggcaggccg cgtcagttgt tgtggccatg 301 gacaagctga ggaagatgct ggttccctgc ccacagacct tccaggagaa tgacctgagc 361 accttctttc ccttcatctt tgaagaagaa cctatcttct tcgacacatg ggataacgag 421 gcttatgtgc acgatgcacc tgtacgatca ctgaactgca cgctccggga ctcacagcaa 481 aaaagcttgg tgatgtctgg tccatatgaa ctgaaagctc tccacctcca gggacaggat 541 atggagcaac aagtggtgtt ctccatgtcc tttgtacaag gagaagaaag taatgacaaa 601 atacctgtgg ccttgggcct caaggaaaag aatctgtacc tgtcctgcgt gttgaaagat 661 gataagccca ctctacagct ggagagtgta gatcccaaaa attacccaaa gaagaagatg 721 gaaaagcgat ttgtcttcaa caagatagaa atcaataaca agctggaatt tgagtctgcc 781 cagttcccca actggtacat cagcacctct caagcagaaa acatgcccgt cttcctggga 841 gggaccaaag gcggccagga tataactgac ttcaccatgc aatttgtgtc ttcctaaaga 901 gagctgtacc cagagagtcc tgtgctgaat gtggactcaa tccctagggc tggcagaaag 961 ggaacagaaa ggtttttgag tacggctata gcctggactt tcctgttgtc tacaccaatg 1021 cccaactgcc tgccttaggg tagtgctaag aggatctcct gtccatcagc caggacagtc 1081 agctctctcc tttcagggcc aatccccagc ccttttgttg agccaggcct ctctcacctc 1141 tcctactcac ttaaagcccg cctgacagaa accacggcca catttggttc taagaaaccc 1201 tctgtcattc gctcccacat tctgatgagc aaccgcttcc ctatttattt atttatttgt 1261 ttgtttgttt tattcattgg tctaatttat tcaaaggggg caagaagtag cagtgtctgt 1321 aaaagagcct agtttttaat agctatggaa tcaattcaat ttggactggt gtgctctctt 1381 taaatcaagt cctttaatta agactgaaaa tatataagct cagattattt aaatgggaat 1441 atttataaat gagcaaatat catactgttc aatggttctg aaataaactt cactgaag Human IL-18 mRNA Variant 1 (SEQ ID NO: 87) 1 attctctccc cagcttgctg agccctttgc tcccctggcg actgcctgga cagtcagcaa 61 ggaattgtct cccagtgcat tttgccctcc tggctgccaa ctctggctgc taaagcggct 121 gccacctgct gcagtctaca cagcttcggg aagaggaaag gaacctcaga ccttccagat 181 cgcttcctct cgcaacaaac tatttgtcgc aggaataaag atggctgctg aaccagtaga 241 agacaattgc atcaactttg tggcaatgaa atttattgac aatacgcttt actttatagc 301 tgaagatgat gaaaacctgg aatcagatta ctttggcaag cttgaatcta aattatcagt 361 cataagaaat ttgaatgacc aagttctctt cattgaccaa ggaaatcggc ctctatttga 421 agatatgact gattctgact gtagagataa tgcaccccgg accatattta ttataagtat 481 gtataaagat agccagccta gaggtatggc tgtaactatc tctgtgaagt gtgagaaaat 541 ttcaactctc tcctgtgaga acaaaattat ttcctttaag gaaatgaatc ctcctgataa 601 catcaaggat acaaaaagtg acatcatatt ctttcagaga agtgtcccag gacatgataa 661 taagatgcaa tttgaatctt catcatacga aggatacttt ctagcttgtg aaaaagagag 721 agacctlitt aaactcattt tgaaaaaaga ggatgaattg ggggatagat ctataatgtt 781 cactgttcaa aacgaagact agctattaaa atttcatgcc gggcgcagtg gctcacgcct 841 gtaatcccag ccctttggga ggctgaggcg ggcagatcac cagaggtcag gtgttcaaga 901 ccagcctgac caacatggtg aaacctcatc tctactaaaa atacaaaaaa ttagctgagt 961 gtagtgacgc atgccctcaa tcccagctac tcaagaggct gaggcaggag aatcacttgc 1021 actccggagg tagaggttgt ggtgagccga gattgcacca ttgcgctcta gcctgggcaa 1081 caacagcaaa actccatctc aaaaaataaa ataaataaat aaacaaataa aaaattcata 1141 atgtgaaaaa aaaaaaaaaa aaa Human IL-18 mRNA Variant 2 (SEQ ID NO: 88) 1 attctctccc cagcttgctg agccctttgc tcccctggcg actgcctgga cagtcagcaa 61 ggaattgtct cccagtgcat tttgccctcc tggctgccaa ctctggctgc taaagcggct 121 gccacctgct gcagtctaca cagcttcggg aagaggaaag gaacctcaga ccttccagat 181 cgcttcctct cgcaacaaac tatttgtcgc aggaataaag atggctgctg aaccagtaga 241 agacaattgc atcaactttg tggcaatgaa atttattgac aatacgcttt actttataga 301 aaacctggaa tcagattact ttggcaagct tgaatctaaa ttatcagtca taagaaattt 361 gaatgaccaa gttctcttca ttgaccaagg aaatcggcct ctatttgaag atatgactga 421 ttctgactgt agagataatg caccccggac catatttatt ataagtatgt ataaagatag 481 ccagcctaga ggtatggctg taactatctc tgtgaagtgt gagaaaattt caactctctc 541 ctgtgagaac aaaattattt cctttaagga aatgaatcct cctgataaca tcaaggatac 601 aaaaagtgac atcatattct ttcagagaag tgtcccagga catgataata agatgcaatt 661 tgaatcttca tcatacgaag gatactttct agcttgtgaa aaagagagag acctttttaa 721 actcattttg aaaaaagagg atgaattggg ggatagatct ataatgttca ctgttcaaaa 781 cgaagactag ctattaaaat ttcatgccgg gcgcagtggc tcacgcctgt aatcccagcc 841 ctttgggagg ctgaggcggg cagatcacca gaggtcaggt gttcaagacc agcctgacca 901 acatggtgaa acctcatctc tactaaaaat acaaaaaatt agctgagtgt agtgacgcat 961 gccctcaatc ccagctactc aagaggctga ggcaggagaa tcacttgcac tccggaggta 1021 gaggttgtgg tgagccgaga ttgcaccatt gcgctctagc ctgggcaaca acagcaaaac 1081 tccatctcaa aaaataaaat aaataaataa acaaataaaa aattcataat gtgaaaaaaa 1141 aaaaaaaaaa a Human IL-36a mRNA (SEQ ID NO: 89) 1 aaaacccaag tgcagtagaa gccattgttc ataatggtag ggatacaggg tccttcgtaa 61 cagattatca gtgtggccta tgctggaaag tctggtgacc tctgattttt tttgcttcca 121 ggtctttggc cttggcactc tttgtcatat tagagttcct gggtctaggc ctgggcagga 181 ttcataggtg cagctgcttc tgctggaggt agactgcatc caacaaagta agggtgctgg 241 gtgagttctg ggagtataga ttctgactgg ggtcactgct gggctggccg ccagtctttc 301 atctgaccca gggttaaact gtggcttggg actgactcag gtcctctctt ggggtcggtc 361 tgcacataaa aggactccta tccttggcag ttctgaaaca acaccaccac aatggaaaaa 421 gcattgaaaa ttgacacacc tcagcagggg agcattcagg atatcaatca tcgggtgtgg 481 gttcttcagg accagacgct catagcagtc ccgaggaagg accgtatgtc tccagtcact 541 attgccttaa tctcatgccg acatgtggag acccttgaga aagacagagg gaaccccatc 601 tacctgggcc tgaatggact caatctctgc ctgatgtgtg ctaaagtcgg ggaccagccc 661 acactgcagc tgaaggaaaa ggatataatg gatttgtaca accaacccga gcctgtgaag 721 tcctttctct tctaccacag ccagagtggc aggaactcca ccttcgagtc tgtggctttc 781 cctggctggt tcatcgctgt cagctctgaa ggaggctgtc ctctcatcct tacccaagaa 841 ctggggaaag ccaacactac tgactttggg ttaactatgc tgttttaa Human IL-36I3 mRNA Variant 1 (SEQ ID NO: 90) 1 cacgggttcc tccccactct gtctttctca cctctccttc acttttccta gcctcctcac 61 caccatctga tctatcttgt tctcttcaca aaaggctctg aagacatcat gaacccacaa 121 cgggaggcag cacccaaatc ctatgctatt cgtgattctc gacagatggt gtgggtcctg 181 agtggaaatt ctttaatagc agctcctctt agccgcagca ttaagcctgt cactcttcat 241 ttaatagcct gtagagacac agaattcagt gacaaggaaa agggtaatat ggtttacctg 301 ggaatcaagg gaaaagatct ctgtctcttc tgtgcagaaa ttcagggcaa gcctactttg 361 cagcttaagc ttcagggctc ccaagataac atagggaagg acacttgctg gaaactagtt 421 ggaattcaca catgcataaa cctggatgtg agagagagct gcttcatggg aacccttgac 481 caatggggaa taggagtggg tagaaagaag tggaagagtt cctttcaaca tcaccatctc 541 aggaagaagg acaaagattt ctcatccatg cggaccaaca taggaatgcc aggaaggatg 601 tagaaataag gggaggaaga ttcccatctc tacaatcttt gagtgggttt gctatcaatg 661 aaatgctaca aatggaataa gttgcagaaa tttttctctt ttcttgggtt ctggagagtt 721 tgtaaaacaa ggacactatg tatttttaaa gagttggtaa atcttacctg taaagctaga 781 gaaggtcgga gtctttttag gagtagattt ggactacata acctgtaaat gtgttttgtc 841 cagtccttag agtgtttttt aaaaaattgt aaagtcaagg ttttcatgaa aaatgggaag 901 atcagacaac attgctcctg aattcccaca gagcagcaag ctactagagc tcaatctgtt 961 atttcttttc ctgatgtaca ggggttaagt cctatggaag aaacagcaga attattcaaa 1021 attatttaca taatgtgcaa ttattcacta gagcatgagg agtgaaacgc tctgtttagt 1081 atgtataact taaaaggaac acatacaatt aaaagtaatt gaaagacatt tcttcttaaa 1141 aattctataa tcttacactg gtaaaataaa ctagtttttc ccatgt Human IL-36I3 mRNA Variant 2 (SEQ ID NO: 91) 1 cacgggttcc tccccactct gtctttctca cctctccttc acttttccta gcctcctcac 61 caccatctga tctatcttgt tctcttcaca aaaggctctg aagacatcat gaacccacaa 121 cgggaggcag cacccaaatc ctatgctatt cgtgattctc gacagatggt gtgggtcctg 181 agtggaaatt ctttaatagc agctcctctt agccgcagca ttaagcctgt cactcttcat 241 ttaatagcct gtagagacac agaattcagt gacaaggaaa agggtaatat ggtttacctg 301 ggaatcaagg gaaaagatct ctgtctcttc tgtgcagaaa ttcagggcaa gcctactttg 361 cagcttaagg aaaaaaatat catggacctg tatgtggaga agaaagcaca gaagcccttt 421 ctctttttcc acaataaaga aggctccact tctgtctttc agtcagtctc ttaccctggc 481 tggttcatag ccacctccac cacatcagga cagcccatct ttctcaccaa ggagagaggc 541 ataactaata acactaactt ctacttagat tctgtggaat aaatccagcc taggctgtgg 601 gtggctggtt ccaggataga gaatcaagct gtcagagtca tcttaacaga tcattatgcg 661 actgagttca ctagcagttc agcccatcca tagcttacct cattcttact atccaaaagc 721 cacctcctcc tccaaacatc catttctgta ccaagaccct cactcgaatg tcactatccc 781 aagatgaaac ctaaaaatca ctttccattc tttcttgatc ttaccccacc atccactcag 841 ctgccatgcc cagtttagtc aaccccccaa atgctgcttc atgcaacctt ccattcctat 901 tccttttgcc aacccatgat gtagagatgt ggattcatga cattttgttc atacaacttc 961 ttcaataaaa cattataata tgtgccccaa agataaagct gaagaatgag atgaatgtga 1021 aattaaaggt ttgcatgtct ttctaatcct aaaaaaaaaa aaaaaaaa Human IL-36y mRNA Variant 1 (SEQ ID NO: 92) 1 gaagctgctg gagccacgat tcagtcccct ggactgtaga taaagaccct ttcttgccag 61 gtgctgagac aaccacacta tgagaggcac tccaggagac gctgatggtg gaggaagggc 121 cgtctatcaa tcaatgtgta aacctattac tgggactatt aatgatttga atcagcaagt 181 gtggaccctt cagggtcaga accttgtggc agttccacga agtgacagtg tgaccccagt 241 cactgttgct gttatcacat gcaagtatcc agaggctctt gagcaaggca gaggggatcc 301 catttatttg ggaatccaga atccagaaat gtgtttgtat tgtgagaagg ttggagaaca 361 gcccacattg cagctaaaag agcagaagat catggatctg tatggccaac ccgagcccgt 421 gaaacccttc cttttctacc gtgccaagac tggtaggacc tccacccttg agtctgtggc 481 cttcccggac tggttcattg cctcctccaa gagagaccag cccatcattc tgacttcaga 541 acttgggaag tcatacaaca ctgcctttga attaaatata aatgactgaa ctcagcctag 601 aggtggcagc ttggtctttg tcttaaagtt tctggttccc aatgtgtttt cgtctacatt 661 ttcttagtgt cattttcacg ctggtgctga gacaggggca aggctgctgt tatcatctca 721 ttttataatg aagaagaagc aattacttca tagcaactga agaacaggat gtggcctcag 781 aagcaggaga gctgggtggt ataaggctgt cctctcaagc tggtgctgtg taggccacaa 841 ggcatctgca tgagtgactt taagactcaa agaccaaaca ctgagctttc ttctaggggt 901 gggtatgaag atgcttcaga gctcatgcgc gttacccacg atggcatgac tagcacagag 961 ctgatctctg tttctgtttt gctttattcc ctcttgggat gatatcatcc agtctttata 1021 tgttgccaat atacctcatt gtgtgtaata gaaccttctt agcattaaga ccttgtaaac 1081 aaaaataatt cttgtgttaa gttaaatcat ttttgtccta attgtaatgt gtaatcttaa 1141 agttaaataa actttgtgta tttatataat aataaagcta aaactgatat aaaataaaga 1201 aagagtaaac tg Human IL-36y mRNA Variant 2 (SEQ ID NO: 93) 1 gaagctgctg gagccacgat tcagtcccct ggactgtaga taaagaccct ttcttgccag 61 gtgctgagac aaccacacta tgagaggcac tccaggagac gctgatggtg gaggaagggc 121 cgtctatcaa tcaatcactg ttgctgttat cacatgcaag tatccagagg ctcttgagca 181 aggcagaggg gatcccattt atttgggaat ccagaatcca gaaatgtgtt tgtattgtga 241 gaaggttgga gaacagccca cattgcagct aaaagagcag aagatcatgg atctgtatgg 301 ccaacccgag cccgtgaaac ccttcctttt ctaccgtgcc aagactggta ggacctccac 361 ccttgagtct gtggccttcc cggactggtt cattgcctcc tccaagagag accagcccat 421 cattctgact tcagaacttg ggaagtcata caacactgcc tttgaattaa atataaatga 481 ctgaactcag cctagaggtg gcagcttggt ctttgtctta aagtttctgg ttcccaatgt 541 gttttcgtct acattttctt agtgtcattt tcacgctggt gctgagacag gggcaaggct 601 gctgttatca tctcatttta taatgaagaa gaagcaatta cttcatagca actgaagaac 661 aggatgtggc ctcagaagca ggagagctgg gtggtataag gctgtcctct caagctggtg 721 ctgtgtaggc cacaaggcat ctgcatgagt gactttaaga ctcaaagacc aaacactgag 781 ctttcttcta ggggtgggta tgaagatgct tcagagctca tgcgcgttac ccacgatggc 841 atgactagca cagagctgat ctctgtttct gttttgcttt attccctctt gggatgatat 901 catccagtct ttatatgttg ccaatatacc tcattgtgtg taatagaacc ttcttagcat 961 taagaccttg taaacaaaaa taattcttgt gttaagttaa atcatttttg tcctaattgt 1021 aatgtgtaat cttaaagtta aataaacttt gtgtatttat ataataataa agctaaaact 1081 gatataaaat aaagaaagag taaactg Human IL-38 mRNA Variant 1 (SEQ ID NO: 94) 1 ggcagtggga ctgggtttga gctgggctta tcctccaact gtgagggagg ctacagcaca 61 ctccacccca ctctcagggc tgggaattgt tgtggctcag ctatttgggg gaatctgttt 121 tccagtttct cagaaccagc gcaagcacac acatcccagg ctcacacccc tggtggctgg 181 acttgctccc ggatagcctc agtcagggag aggcagagct gcctggagcc tgctgggctg 241 gtggaagcct tggtggattc tggcaggcca attatagacg aatggcctgg ggaacccgtg 301 cagcccttgg ctgagtggtt ctaagcccca gcacgtctgc ctctggcttc acccagcctc 361 cttttctaac tgcccttctc tcctccccat cagtgaggac cagacaccac tgattgcagg 421 aatgtgttcc ctccccatgg caagatacta cataattaaa tatgcagacc agaaggctct 481 atacacaaga gatggccagc tgctggtggg agatcctgtt gcagacaact gctgtgcaga 541 gaagatctgc atacttccta acagaggctt ggcccgcacc aaggtcccca ttttcctggg 601 gatccaggga gggagccgct gcctggcatg tgtggagaca gaagaggggc cttccctaca 661 gctggaggat gtgaacattg aggaactgta caaaggtggt gaagaggcca cacgcttcac 721 cttcttccag agcagctcag gctccgcctt caggcttgag gctgctgcct ggcctggctg 781 gttcctgtgt ggcccggcag agccccagca gccagtacag ctcaccaagg agagtgagcc 841 ctcagcccgt accaagtttt actttgaaca gagctggtag ggagacagga aactgcgttt 901 tagccttgtg cccccaaacc aagctcatcc tgctcagggt ctatggtagg cagaataatg 961 tcccccgaaa tatgtccaca tcctaatccc aagatctgtg catatgttac catacatgtc 1021 caaagaggtt ttgcaaatgt gattatgtta aggatcttga aatgaggaga caatcctggg 1081 ttatccttgt gggctcagtt taatcacaag aaggaggcag gaagggagag tcagagagag 1141 aatggaagat accatgcttc taattttgaa gatggagtga ggggccttga gccaacaaat 1201 gcaggtgttt ttagaaggtg gaaaagccaa gggaacggat tctcctctag agtctccgga 1261 aggaacacag ctcttgacac atggatttca gctcagtgac acccatttca gacttctgac 1321 ctccacaact ataaaataat aaacttgtgt tattgtaaac ctctaa Human IL-38 mRNA Variant 2 (SEQ ID NO: 95) 1 agttggagtc tccagggatc agggttccag gaactcagga tctgcagtga ggaccagaca 61 ccactgattg caggaatgtg ttccctcccc atggcaagat actacataat taaatatgca 121 gaccagaagg ctctatacac aagagatggc cagctgctgg tgggagatcc tgttgcagac 181 aactgctgtg cagagaagat ctgcatactt cctaacagag gcttggcccg caccaaggtc 241 cccattttcc tggggatcca gggagggagc cgctgcctgg catgtgtgga gacagaagag 301 gggccttccc tacagctgga ggatgtgaac attgaggaac tgtacaaagg tggtgaagag 361 gccacacgct tcaccttctt ccagagcagc tcaggctccg ccttcaggct tgaggctgct 421 gcctggcctg gctggttcct gtgtggcccg gcagagcccc agcagccagt acagctcacc 481 aaggagagtg agccctcagc ccgtaccaag ttttactttg aacagagctg gtagggagac 541 aggaaactgc gttttagcct tgtgccccca aaccaagctc atcctgctca gggtctatgg 601 taggcagaat aatgtccccc gaaatatgtc cacatcctaa tcccaagatc tgtgcatatg 661 ttaccataca tgtccaaaga ggttttgcaa atgtgattat gttaaggatc ttgaaatgag 721 gagacaatcc tgggttatcc ttgtgggctc agtttaatca caagaaggag gcaggaaggg 781 agagtcagag agagaatgga agataccatg cttctaattt tgaagatgga gtgaggggcc 841 ttgagccaac aaatgcaggt gtttttagaa ggtggaaaag ccaagggaac ggattctcct 901 ctagagtctc cggaaggaac acagctcttg acacatggat ttcagctcag tgacacccat 961 ttcagacttc tgacctccac aactataaaa taataaactt gtgttattgt aaacctctaa 1021 aaaaaaa Human IL-33 mRNA Variant 1 (SEQ ID NO: 96) 1 agtctacaga ctcctccgaa cacagagctg cagctcttca gggaagaaat caaaacaaga 61 tcacaagaat actgaaaaat gaagcctaaa atgaagtatt caaccaacaa aatttccaca 121 gcaaagtgga agaacacagc aagcaaagcc ttgtgtttca agctgggaaa atcccaacag 181 aaggccaaag aagtttgccc catgtacttt atgaagctcc gctctggcct tatgataaaa 241 aaggaggcct gttactttag gagagaaacc accaaaaggc cttcactgaa aacaggtaga 301 aagcacaaaa gacatctggt actcgctgcc tgtcaacagc agtctactgt ggagtgcttt 361 gcctttggta tatcaggggt ccagaaatat actagagcac ttcatgattc aagtatcaca 421 ggaatttcac ctattacaga gtatcttgct tctctaagca catacaatga tcaatccatt 481 acttttgctt tggaggatga aagttatgag atatatgttg aagacttgaa aaaagatgaa 541 aagaaagata aggtgttact gagttactat gagtctcaac acccctcaaa tgaatcaggt 601 gacggtgttg atggtaagat gttaatggta accctgagtc ctacaaaaga cttctggttg 661 catgccaaca acaaggaaca ctctgtggag ctccataagt gtgaaaaacc actgccagac 721 caggccttct ttgtccttca taatatgcac tccaactgtg tttcatttga atgcaagact 781 gatcctggag tgtttatagg tgtaaaggat aatcatcttg ctctgattaa agtagactct 841 tctgagaatt tgtgtactga aaatatcttg tttaagctct ctgaaactta gttgatggaa 901 acctgtgagt cttgggttga gtacccaaat gctaccactg gagaaggaat gagagataaa 961 gaaagagaca ggtgacatct aagggaaatg aagagtgctt agcatgtgtg gaatgttttc 1021 catattatgt ataaaaatat tttttctaat cctccagtta ttcttttatt tccctctgta 1081 taactgcatc ttcaatacaa gtatcagtat attaaatagg gtattggtaa agaaacggtc 1141 aacattctaa agagatacag tctgaccttt acttttctct agtttcagtc cagaaagaac 1201 ttcatattta gagctaaggc cactgaggaa agagccatag cttaagtctc tatgtagaca 1261 gggatccatt ttaaagagct acttagagaa ataattttcc acagttccaa acgataggct 1321 caaacactag agctgctagt aaaaagaaga ccagatgctt cacagaatta tcattttttc 1381 aactggaata aaacaccagg tttgtttgta gatgtcttag gcaacactca gagcagatct 1441 cccttactgt caggggatat ggaacttcaa aggcccacat ggcaagccag gtaacataaa 1501 tgtgtgaaaa agtaaagata actaaaaaat ttagaaaaat aaatccagta tttgtaaagt 1561 gaataacttc atttctaatt gtttaatttt taaaattctg atttttatat attgagttta 1621 agcaaggcat tcttacacga ggaagtgaag taaattttag ttcagacata aaatttcact 1681 tattaggaat atgtaacatg ctaaaacttt ttttttttta aagagtactg agtcacaaca 1741 tgttttagag catccaagta ccatataatc caactatcat ggtaaggcca gaaatcttct 1801 aacctaccag agcctagatg agacaccgaa ttaacattaa aatttcagta actgactgtc 1861 cctcatgtcc atggcctacc atcccttctg accctggctt ccagggacct atgtctttta 1921 atactcactg tcacattggg caaagttgct tctaatcctt atttcccatg tgcacaagtc 1981 tttttgtatt ccagcttcct gataacactg cttactgtgg aatattcatt tgacatctgt 2041 ctcttttcat ttcttttaac taccatgccc ttgatatatc ttttgcacct gctgaacttc 2101 atttctgtat cacctgacct ctggatgcca aaacgtttat tctgctttgt ctgttgtaga 2161 attttagata aagctattaa tggcaatatt tttttgctaa acgtttttgt tttttactgt 2221 cactagggca ataaaattta tactcaacca tataataaca ttttttaact actaaaggag 2281 tagtttttat tttaaagtct tagcaatttc tattacaact tttcttagac ttaacactta 2341 tgataaatga ctaacatagt aacagaatct ttatgaaata tgaccttttc tgaaaataca 2401 tacttttaca tttctacttt attgagacct attagatgta agtgctagta gaatataaga 2461 taaaagaggc tgagaattac catacaaggg tattacaact gtaaaacaat ttatctttgt 2521 ttcattgttc tgtcaataat tgttaccaaa gagataaaaa taaaagcaga atgtatatca 2581 tcccatctga aaaacactaa ttattgacat gtgcatctgt acaataaact taaaatgatt 2641 attaaataat caaatatatc tactacattg tttatattat tgaataaagt atattttcca 2701 aatgtaaaaa aaaaaaaa Human IL-33 mRNA Variant 2 (SEQ ID NO: 97) 1 agtctacaga ctcctccgaa cacagagctg cagctcttca gggaagaaat caaaacaaga 61 tcacaagaat actgaaaaat gaagcctaaa atgaagtatt caaccaacaa aatttccaca 121 gcaaagtgga agaacacagc aagcaaagcc ttgtgtttca agctgggaaa atcccaacag 181 aaggccaaag aagtttgccc catgtacttt atgaagctcc gctctggcct tatgataaaa 241 aaggaggcct gttactttag gagagaaacc accaaaaggc cttcactgaa aacaggtaga 301 aagcacaaaa gacatctggt actcgctgcc tgtcaacagc agtctactgt ggagtgcttt 361 gcctttggta tatcaggggt ccagaaatat actagagcac ttcatgattc aagtatcaca 421 gataaggtgt tactgagtta ctatgagtct caacacccct caaatgaatc aggtgacggt 481 gttgatggta agatgttaat ggtaaccctg agtcctacaa aagacttctg gttgcatgcc 541 aacaacaagg aacactctgt ggagctccat aagtgtgaaa aaccactgcc agaccaggcc 601 ttctttgtcc ttcataatat gcactccaac tgtgtttcat ttgaatgcaa gactgatcct 661 ggagtgttta taggtgtaaa ggataatcat cttgctctga ttaaagtaga ctcttctgag 721 aatttgtgta ctgaaaatat cttgtttaag ctctctgaaa cttagttgat ggaaacctgt 781 gagtcttggg ttgagtaccc aaatgctacc actggagaag gaatgagaga taaagaaaga 841 gacaggtgac atctaaggga aatgaagagt gcttagcatg tgtggaatgt tttccatatt 901 atgtataaaa atattttttc taatcctcca gttattcttt tatttccctc tgtataactg 961 catcttcaat acaagtatca gtatattaaa tagggtattg gtaaagaaac ggtcaacatt 1021 ctaaagagat acagtctgac ctttactttt ctctagtttc agtccagaaa gaacttcata 1081 tttagagcta aggccactga ggaaagagcc atagcttaag tctctatgta gacagggatc 1141 cattttaaag agctacttag agaaataatt ttccacagtt ccaaacgata ggctcaaaca 1201 ctagagctgc tagtaaaaag aagaccagat gcttcacaga attatcattt tttcaactgg 1261 aataaaacac caggtttgtt tgtagatgtc ttaggcaaca ctcagagcag atctccctta 1321 ctgtcagggg atatggaact tcaaaggccc acatggcaag ccaggtaaca taaatgtgtg 1381 aaaaagtaaa gataactaaa aaatttagaa aaataaatcc agtatttgta aagtgaataa 1441 cttcatttct aattgtttaa tttttaaaat tctgattttt atatattgag tttaagcaag 1501 gcattcttac acgaggaagt gaagtaaatt ttagttcaga cataaaattt cacttattag 1561 gaatatgtaa catgctaaaa cttttttttt tttaaagagt actgagtcac aacatgtttt 1621 agagcatcca agtaccatat aatccaacta tcatggtaag gccagaaatc ttctaaccta 1681 ccagagccta gatgagacac cgaattaaca ttaaaatttc agtaactgac tgtccctcat 1741 gtccatggcc taccatccct tctgaccctg gcttccaggg acctatgtct tttaatactc 1801 actgtcacat tgggcaaagt tgcttctaat ccttatttcc catgtgcaca agtctttttg 1861 tattccagct tcctgataac actgcttact gtggaatatt catttgacat ctgtctcttt 1921 tcatttcttt taactaccat gcccttgata tatcttttgc acctgctgaa cttcatttct 1981 gtatcacctg acctctggat gccaaaacgt ttattctgct ttgtctgttg tagaatttta 2041 gataaagcta ttaatggcaa tatttttttg ctaaacgttt ttgtttttta ctgtcactag 2101 ggcaataaaa tttatactca accatataat aacatttttt aactactaaa ggagtagttt 2161 ttattttaaa gtcttagcaa tttctattac aacttttctt agacttaaca cttatgataa 2221 atgactaaca tagtaacaga atctttatga aatatgacct tttctgaaaa tacatacttt 2281 tacatttcta ctttattgag acctattaga tgtaagtgct agtagaatat aagataaaag 2341 aggctgagaa ttaccataca agggtattac aactgtaaaa caatttatct ttgtttcatt 2401 gttctgtcaa taattgttac caaagagata aaaataaaag cagaatgtat atcatcccat 2461 ctgaaaaaca ctaattattg acatgtgcat ctgtacaata aacttaaaat gattattaaa 2521 taatcaaata tatctactac attgtttata ttattgaata aagtatattt tccaaatgta 2581 aaaaaaaaaa aa Human IL-33 mRNA Variant 3 (SEQ ID NO: 98) 1 agtctacaga ctcctccgaa cacagagctg cagctcttca gggaagaaat caaaacaaga 61 tcacaagaat actgaaaaat gaagcctaaa atgaagtatt caaccaacaa aatttccaca 121 gcaaagtgga agaacacagc aagcaaagcc ttgtgtttca agctgggaaa taaggtgtta 181 ctgagttact atgagtctca acacccctca aatgaatcag gtgacggtgt tgatggtaag 241 atgttaatgg taaccctgag tcctacaaaa gacttctggt tgcatgccaa caacaaggaa 301 cactctgtgg agctccataa gtgtgaaaaa ccactgccag accaggcctt ctttgtcctt 361 cataatatgc actccaactg tgtttcattt gaatgcaaga ctgatcctgg agtgtttata 421 ggtgtaaagg ataatcatct tgctctgatt aaagtagact cttctgagaa tttgtgtact 481 gaaaatatct tgtttaagct ctctgaaact tagttgatgg aaacctgtga gtcttgggtt 541 gagtacccaa atgctaccac tggagaagga atgagagata aagaaagaga caggtgacat 601 ctaagggaaa tgaagagtgc ttagcatgtg tggaatgttt tccatattat gtataaaaat 661 attttttcta atcctccagt tattctttta tttccctctg tataactgca tcttcaatac 721 aagtatcagt atattaaata gggtattggt aaagaaacgg tcaacattct aaagagatac 781 agtctgacct ttacttttct ctagtttcag tccagaaaga acttcatatt tagagctaag 841 gccactgagg aaagagccat agcttaagtc tctatgtaga cagggatcca ttttaaagag 901 ctacttagag aaataatttt ccacagttcc aaacgatagg ctcaaacact agagctgcta 961 gtaaaaagaa gaccagatgc ttcacagaat tatcattttt tcaactggaa taaaacacca 1021 ggtttgtttg tagatgtctt aggcaacact cagagcagat ctcccttact gtcaggggat 1081 atggaacttc aaaggcccac atggcaagcc aggtaacata aatgtgtgaa aaagtaaaga 1141 taactaaaaa atttagaaaa ataaatccag tatttgtaaa gtgaataact tcatttctaa 1201 ttgtttaatt tttaaaattc tgatttttat atattgagtt taagcaaggc attcttacac 1261 gaggaagtga agtaaatttt agttcagaca taaaatttca cttattagga atatgtaaca 1321 tgctaaaact tttttttttt taaagagtac tgagtcacaa catgttttag agcatccaag 1381 taccatataa tccaactatc atggtaaggc cagaaatctt ctaacctacc agagcctaga 1441 tgagacaccg aattaacatt aaaatttcag taactgactg tccctcatgt ccatggccta 1501 ccatcccttc tgaccctggc ttccagggac ctatgtcttt taatactcac tgtcacattg 1561 ggcaaagttg cttctaatcc ttatttccca tgtgcacaag tctttttgta ttccagcttc 1621 ctgataacac tgcttactgt ggaatattca tttgacatct gtctcttttc atttctttta 1681 actaccatgc ccttgatata tcttttgcac ctgctgaact tcatttctgt atcacctgac 1741 ctctggatgc caaaacgttt attctgcttt gtctgttgta gaattttaga taaagctatt 1801 aatggcaata tttttttgct aaacgttttt gttttttact gtcactaggg caataaaatt 1861 tatactcaac catataataa cattttttaa ctactaaagg agtagttttt attttaaagt 1921 cttagcaatt tctattacaa cttttcttag acttaacact tatgataaat gactaacata 1981 gtaacagaat ctttatgaaa tatgaccttt tctgaaaata catactttta catttctact 2041 ttattgagac ctattagatg taagtgctag tagaatataa gataaaagag gctgagaatt 2101 accatacaag ggtattacaa ctgtaaaaca atttatcttt gtttcattgt tctgtcaata 2161 attgttacca aagagataaa aataaaagca gaatgtatat catcccatct gaaaaacact 2221 aattattgac atgtgcatct gtacaataaa cttaaaatga ttattaaata atcaaatata 2281 tctactacat tgtttatatt attgaataaa gtatattttc caaatgtaaa aaaaaaaaaa Human IL-33 mRNA Variant 4 (SEQ ID NO: 99) 1 acagatgcca aacgagatgg agagagggtg agtaggagca aaatttctca tgagaatact 61 gaaaaatgaa gcctaaaatg aagtattcaa ccaacaaaat ttccacagca aagtggaaga 121 acacagcaag caaagccttg tgtttcaagc tgggaaaatc ccaacagaag gccaaagaag 181 tttgccccat gtactttatg aagctccgct ctggccttat gataaaaaag gaggcctgtt 241 actttaggag agaaaccacc aaaaggcctt cactgaaaac aggtagaaag cacaaaagac 301 atctggtact cgctgcctgt caacagcagt ctactgtgga gtgctttgcc tttggtatat 361 caggggtcca gaaatatact agagcacttc atgattcaag tatcacagga atttcaccta 421 ttacagagta tcttgcttct ctaagcacat acaatgatca atccattact tttgctttgg 481 aggatgaaag ttatgagata tatgttgaag acttgaaaaa agatgaaaag aaagataagg 541 tgttactgag ttactatgag tctcaacacc cctcaaatga atcaggtgac ggtgttgatg 601 gtaagatgtt aatggtaacc ctgagtccta caaaagactt ctggttgcat gccaacaaca 661 aggaacactc tgtggagctc cataagtgtg aaaaaccact gccagaccag gccttctttg 721 tccttcataa tatgcactcc aactgtgttt catttgaatg caagactgat cctggagtgt 781 ttataggtgt aaaggataat catcttgctc tgattaaagt agactcttct gagaatttgt 841 gtactgaaaa tatcttgttt aagctctctg aaacttagtt gatggaaacc tgtgagtctt 901 gggttgagta cccaaatgct accactggag aaggaatgag agataaagaa agagacaggt 961 gacatctaag ggaaatgaag agtgcttagc atgtgtggaa tgttttccat attatgtata 1021 aaaatatttt ttctaatcct ccagttattc ttttatttcc ctctgtataa ctgcatcttc 1081 aatacaagta tcagtatatt aaatagggta ttggtaaaga aacggtcaac attctaaaga 1141 gatacagtct gacctttact tttctctagt ttcagtccag aaagaacttc atatttagag 1201 ctaaggccac tgaggaaaga gccatagctt aagtctctat gtagacaggg atccatttta 1261 aagagctact tagagaaata attttccaca gttccaaacg ataggctcaa acactagagc 1321 tgctagtaaa aagaagacca gatgcttcac agaattatca ttttttcaac tggaataaaa 1381 caccaggttt gtttgtagat gtcttaggca acactcagag cagatctccc ttactgtcag 1441 gggatatgga acttcaaagg cccacatggc aagccaggta acataaatgt gtgaaaaagt 1501 aaagataact aaaaaattta gaaaaataaa tccagtattt gtaaagtgaa taacttcatt 1561 tctaattgtt taatttttaa aattctgatt tttatatatt gagtttaagc aaggcattct 1621 tacacgagga agtgaagtaa attttagttc agacataaaa tttcacttat taggaatatg 1681 taacatgcta aaactttttt ttttttaaag agtactgagt cacaacatgt tttagagcat 1741 ccaagtacca tataatccaa ctatcatggt aaggccagaa atcttctaac ctaccagagc 1801 ctagatgaga caccgaatta acattaaaat ttcagtaact gactgtccct catgtccatg 1861 gcctaccatc ccttctgacc ctggcttcca gggacctatg tcttttaata ctcactgtca 1921 cattgggcaa agttgcttct aatccttatt tcccatgtgc acaagtcttt ttgtattcca 1981 gcttcctgat aacactgctt actgtggaat attcatttga catctgtctc ttttcatttc 2041 ttttaactac catgcccttg atatatcttt tgcacctgct gaacttcatt tctgtatcac 2101 ctgacctctg gatgccaaaa cgtttattct gctttgtctg ttgtagaatt ttagataaag 2161 ctattaatgg caatattttt ttgctaaacg tttttgtttt ttactgtcac tagggcaata 2221 aaatttatac tcaaccatat aataacattt tttaactact aaaggagtag tttttatttt 2281 aaagtcttag caatttctat tacaactttt cttagactta acacttatga taaatgacta 2341 acatagtaac agaatcttta tgaaatatga ccttttctga aaatacatac ttttacattt 2401 ctactttatt gagacctatt agatgtaagt gctagtagaa tataagataa aagaggctga 2461 gaattaccat acaagggtat tacaactgta aaacaattta tctttgtttc attgttctgt 2521 caataattgt taccaaagag ataaaaataa aagcagaatg tatatcatcc catctgaaaa 2581 acactaatta ttgacatgtg catctgtaca ataaacttaa aatgattatt aaataatcaa 2641 atatatctac tacattgttt atattattga ataaagtata ttttccaaat gtaaaaaaaa 2701 aaaaa Human IL-33 mRNA Variant 5 (SEQ ID NO: 100) 1 aaatactaca attgctgact acaggaaacc tcatcatctg agaccagcac tttataaatt 61 agaatactga aaaatgaagc ctaaaatgaa gtattcaacc aacaaaattt ccacagcaaa 121 gtggaagaac acagcaagca aagccttgtg tttcaagctg ggaaaatccc aacagaaggc 181 caaagaagtt tgccccatgt actttatgaa gctccgctct ggccttatga taaaaaagga 241 ggcctgttac tttaggagag aaaccaccaa aaggccttca ctgaaaacag gtagaaagca 301 caaaagacat ctggtactcg ctgcctgtca acagcagtct actgtggagt gctttgcctt 361 tggtatatca ggggtccaga aatatactag agcacttcat gattcaagta tcacaggaat 421 ttcacctatt acagagtatc ttgcttctct aagcacatac aatgatcaat ccattacttt 481 tgctttggag gatgaaagtt atgagatata tgttgaagac ttgaaaaaag atgaaaagaa 541 agataaggtg ttactgagtt actatgagtc tcaacacccc tcaaatgaat caggtgacgg 601 tgttgatggt aagatgttaa tggtaaccct gagtcctaca aaagacttct ggttgcatgc 661 caacaacaag gaacactctg tggagctcca taagtgtgaa aaaccactgc cagaccaggc 721 cttctttgtc cttcataata tgcactccaa ctgtgtttca tttgaatgca agactgatcc 781 tggagtgttt ataggtgtaa aggataatca tcttgctctg attaaagtag actcttctga 841 gaatttgtgt actgaaaata tcttgtttaa gctctctgaa acttagttga tggaaacctg 901 tgagtcttgg gttgagtacc caaatgctac cactggagaa ggaatgagag ataaagaaag 961 agacaggtga catctaaggg aaatgaagag tgcttagcat gtgtggaatg ttttccatat 1021 tatgtataaa aatatttttt ctaatcctcc agttattctt ttatttccct ctgtataact 1081 gcatcttcaa tacaagtatc agtatattaa atagggtatt ggtaaagaaa cggtcaacat 1141 tctaaagaga tacagtctga cctttacttt tctctagttt cagtccagaa agaacttcat 1201 atttagagct aaggccactg aggaaagagc catagcttaa gtctctatgt agacagggat 1261 ccattttaaa gagctactta gagaaataat tttccacagt tccaaacgat aggctcaaac 1321 actagagctg ctagtaaaaa gaagaccaga tgcttcacag aattatcatt ttttcaactg 1381 gaataaaaca ccaggtttgt ttgtagatgt cttaggcaac actcagagca gatctccctt 1441 actgtcaggg gatatggaac ttcaaaggcc cacatggcaa gccaggtaac ataaatgtgt 1501 gaaaaagtaa agataactaa aaaatttaga aaaataaatc cagtatttgt aaagtgaata 1561 acttcatttc taattgttta atttttaaaa ttctgatttt tatatattga gtttaagcaa 1621 ggcattctta cacgaggaag tgaagtaaat tttagttcag acataaaatt tcacttatta 1681 ggaatatgta acatgctaaa actttttttt ttttaaagag tactgagtca caacatgttt 1741 tagagcatcc aagtaccata taatccaact atcatggtaa ggccagaaat cttctaacct 1801 accagagcct agatgagaca ccgaattaac attaaaattt cagtaactga ctgtccctca 1861 tgtccatggc ctaccatccc ttctgaccct ggcttccagg gacctatgtc ttttaatact 1921 cactgtcaca ttgggcaaag ttgcttctaa tccttatttc ccatgtgcac aagtcttttt 1981 gtattccagc ttcctgataa cactgcttac tgtggaatat tcatttgaca tctgtctctt 2041 ttcatttctt ttaactacca tgcccttgat atatcttttg cacctgctga acttcatttc 2101 tgtatcacct gacctctgga tgccaaaacg tttattctgc tttgtctgtt gtagaatttt 2161 agataaagct attaatggca atattttttt gctaaacgtt tttgtttttt actgtcacta 2221 gggcaataaa atttatactc aaccatataa taacattttt taactactaa aggagtagtt 2281 tttattttaa agtcttagca atttctatta caacttttct tagacttaac acttatgata 2341 aatgactaac atagtaacag aatctttatg aaatatgacc ttttctgaaa atacatactt 2401 ttacatttct actttattga gacctattag atgtaagtgc tagtagaata taagataaaa 2461 gaggctgaga attaccatac aagggtatta caactgtaaa acaatttatc tttgtttcat 2521 tgttctgtca ataattgtta ccaaagagat aaaaataaaa gcagaatgta tatcatccca 2581 tctgaaaaac actaattatt gacatgtgca tctgtacaat aaacttaaaa tgattattaa 2641 ataatcaaat atatctacta cattgtttat attattgaat aaagtatatt ttccaaatgt 2701 aaaaaaaaaa aaa Human IL-33 mRNA Variant 6 (SEQ ID NO: 101) 1 agtctacaga ctcctccgaa cacagagctg cagctcttca gggaagaaat caaaacaaga 61 tcacaagaat actgaaaaat gaagcctaaa atgaagtatt caaccaacaa aatttccaca 121 gcaaagtgga agaacacagc aagcaaagcc ttgtgtttca agctgggaaa atcccaacag 181 aaggccaaag aagtttgccc catgtacttt atgaagctcc gctctggcct tatgataaaa 241 aaggaggcct gttactttag gagagaaacc accaaaaggc cttcactgaa aacaggtaga 301 aagcacaaaa gacatctggt actcgctgcc tgtcaacagc agtctactgt ggagtgcttt 361 gcctttggta tatcaggggt ccagaaatat actagagcac ttcatgattc aagtatcaca 421 gagtatcttg cttctctaag cacatacaat gatcaatcca ttacttttgc tttggaggat 481 gaaagttatg agatatatgt tgaagacttg aaaaaagatg aaaagaaaga taaggtgtta 541 ctgagttact atgagtctca acacccctca aatgaatcag gtgacggtgt tgatggtaag 601 atgttaatgg taaccctgag tcctacaaaa gacttctggt tgcatgccaa caacaaggaa 661 cactctgtgg agctccataa gtgtgaaaaa ccactgccag accaggcctt ctttgtcctt 721 cataatatgc actccaactg tgtttcattt gaatgcaaga ctgatcctgg agtgtttata 781 ggtgtaaagg ataatcatct tgctctgatt aaagtagact cttctgagaa tttgtgtact 841 gaaaatatct tgtttaagct ctctgaaact tagttgatgg aaacctgtga gtcttgggtt 901 gagtacccaa atgctaccac tggagaagga atgagagata aagaaagaga caggtgacat 961 ctaagggaaa tgaagagtgc ttagcatgtg tggaatgttt tccatattat gtataaaaat 1021 attttttcta atcctccagt tattctttta tttccctctg tataactgca tcttcaatac 1081 aagtatcagt atattaaata gggtattggt aaagaaacgg tcaacattct aaagagatac 1141 agtctgacct ttacttttct ctagtttcag tccagaaaga acttcatatt tagagctaag 1201 gccactgagg aaagagccat agcttaagtc tctatgtaga cagggatcca ttttaaagag 1261 ctacttagag aaataatttt ccacagttcc aaacgatagg ctcaaacact agagctgcta 1321 gtaaaaagaa gaccagatgc ttcacagaat tatcattttt tcaactggaa taaaacacca 1381 ggtttgtttg tagatgtctt aggcaacact cagagcagat ctcccttact gtcaggggat 1441 atggaacttc aaaggcccac atggcaagcc aggtaacata aatgtgtgaa aaagtaaaga 1501 taactaaaaa atttagaaaa ataaatccag tatttgtaaa gtgaataact tcatttctaa 1561 ttgtttaatt tttaaaattc tgatttttat atattgagtt taagcaaggc attcttacac 1621 gaggaagtga agtaaatttt agttcagaca taaaatttca cttattagga atatgtaaca 1681 tgctaaaact tttttttttt taaagagtac tgagtcacaa catgttttag agcatccaag 1741 taccatataa tccaactatc atggtaaggc cagaaatctt ctaacctacc agagcctaga 1801 tgagacaccg aattaacatt aaaatttcag taactgactg tccctcatgt ccatggccta 1861 ccatcccttc tgaccctggc ttccagggac ctatgtcttt taatactcac tgtcacattg 1921 ggcaaagttg cttctaatcc ttatttccca tgtgcacaag tctttttgta ttccagcttc 1981 ctgataacac tgcttactgt ggaatattca tttgacatct gtctcttttc atttctttta 2041 actaccatgc ccttgatata tcttttgcac ctgctgaact tcatttctgt atcacctgac 2101 ctctggatgc caaaacgttt attctgcttt gtctgttgta gaattttaga taaagctatt 2161 aatggcaata tttttttgct aaacgttttt gttttttact gtcactaggg caataaaatt 2221 tatactcaac catataataa cattttttaa ctactaaagg agtagttttt attttaaagt 2281 cttagcaatt tctattacaa cttttcttag acttaacact tatgataaat gactaacata 2341 gtaacagaat ctttatgaaa tatgaccttt tctgaaaata catactttta catttctact 2401 ttattgagac ctattagatg taagtgctag tagaatataa gataaaagag gctgagaatt 2461 accatacaag ggtattacaa ctgtaaaaca atttatcttt gtttcattgt tctgtcaata 2521 attgttacca aagagataaa aataaaagca gaatgtatat catcccatct gaaaaacact 2581 aattattgac atgtgcatct gtacaataaa cttaaaatga ttattaaata atcaaatata 2641 tctactacat tgtttatatt attgaataaa gtatattttc caaatgtaaa aaaaaaaaaa Human IL-33 mRNA Variant 7 (SEQ ID NO: 102) 1 acagatgcca aacgagatgg agagagggtg agtaggagca aaatttctca tgagaatact 61 gaaaaatgaa gcctaaaatg aagtattcaa ccaacaaaat ttccacagca aagtggaaga 121 acacagcaag caaagccttg tgtttcaagc tgggaaaatc ccaacagaag gccaaagaag 181 tttgccccat gtactttatg aagctccgct ctggccttat gataaaaaag gaggcctgtt 241 actttaggag agaaaccacc aaaaggcctt cactgaaaac aggtagaaag cacaaaagac 301 atctggtact cgctgcctgt caacagcagt ctactgtgga gtgctttgcc tttggtatat 361 caggggtcca gaaatatact agagcacttc atgattcaag tatcacagag tatcttgctt 421 ctctaagcac atacaatgat caatccatta cttttgcttt ggaggatgaa agttatgaga 481 tatatgttga agacttgaaa aaagatgaaa agaaagataa ggtgttactg agttactatg 541 agtctcaaca cccctcaaat gaatcaggtg acggtgttga tggtaagatg ttaatggtaa 601 ccctgagtcc tacaaaagac ttctggttgc atgccaacaa caaggaacac tctgtggagc 661 tccataagtg tgaaaaacca ctgccagacc aggccttctt tgtccttcat aatatgcact 721 ccaactgtgt ttcatttgaa tgcaagactg atcctggagt gtttataggt gtaaaggata 781 atcatcttgc tctgattaaa gtagactctt ctgagaattt gtgtactgaa aatatcttgt 841 ttaagctctc tgaaacttag ttgatggaaa cctgtgagtc ttgggttgag tacccaaatg 901 ctaccactgg agaaggaatg agagataaag aaagagacag gtgacatcta agggaaatga 961 agagtgctta gcatgtgtgg aatgttttcc atattatgta taaaaatatt ttttctaatc 1021 ctccagttat tcttttattt ccctctgtat aactgcatct tcaatacaag tatcagtata 1081 ttaaataggg tattggtaaa gaaacggtca acattctaaa gagatacagt ctgaccttta 1141 cttttctcta gtttcagtcc agaaagaact tcatatttag agctaaggcc actgaggaaa 1201 gagccatagc ttaagtctct atgtagacag ggatccattt taaagagcta cttagagaaa 1261 taattttcca cagttccaaa cgataggctc aaacactaga gctgctagta aaaagaagac 1321 cagatgcttc acagaattat cattttttca actggaataa aacaccaggt ttgtttgtag 1381 atgtcttagg caacactcag agcagatctc ccttactgtc aggggatatg gaacttcaaa 1441 ggcccacatg gcaagccagg taacataaat gtgtgaaaaa gtaaagataa ctaaaaaatt 1501 tagaaaaata aatccagtat ttgtaaagtg aataacttca tttctaattg tttaattttt 1561 aaaattctga tttttatata ttgagtttaa gcaaggcatt cttacacgag gaagtgaagt 1621 aaattttagt tcagacataa aatttcactt attaggaata tgtaacatgc taaaactttt 1681 ttttttttaa agagtactga gtcacaacat gttttagagc atccaagtac catataatcc 1741 aactatcatg gtaaggccag aaatcttcta acctaccaga gcctagatga gacaccgaat 1801 taacattaaa atttcagtaa ctgactgtcc ctcatgtcca tggcctacca tcccttctga 1861 ccctggcttc cagggaccta tgtcttttaa tactcactgt cacattgggc aaagttgctt 1921 ctaatcctta tttcccatgt gcacaagtct ttttgtattc cagcttcctg ataacactgc 1981 ttactgtgga atattcattt gacatctgtc tcttttcatt tcttttaact accatgccct 2041 tgatatatct tttgcacctg ctgaacttca tttctgtatc acctgacctc tggatgccaa 2101 aacgtttatt ctgctttgtc tgttgtagaa ttttagataa agctattaat ggcaatattt 2161 ttttgctaaa cgtttttgtt ttttactgtc actagggcaa taaaatttat actcaaccat 2221 ataataacat tttttaacta ctaaaggagt agtttttatt ttaaagtctt agcaatttct 2281 attacaactt ttcttagact taacacttat gataaatgac taacatagta acagaatctt 2341 tatgaaatat gaccttttct gaaaatacat acttttacat ttctacttta ttgagaccta 2401 ttagatgtaa gtgctagtag aatataagat aaaagaggct gagaattacc atacaagggt 2461 attacaactg taaaacaatt tatctttgtt tcattgttct gtcaataatt gttaccaaag 2521 agataaaaat aaaagcagaa tgtatatcat cccatctgaa aaacactaat tattgacatg 2581 tgcatctgta caataaactt aaaatgatta ttaaataatc aaatatatct actacattgt 2641 ttatattatt gaataaagta tattttccaa atgtaaaaaa aaaaaaa Human IL-33 mRNA Variant 8 (SEQ ID NO: 103) 1 agtctacaga ctcctccgaa cacagagctg cagctcttca gggaagaaat caaaacaaga 61 tcacaagaat actgaaaaat gaagcctaaa atgaagtatt caaccaacaa aatttccaca 121 gcaaagtgga agaacacagc aagcaaagcc ttgtgtttca agctgggaaa atcccaacag 181 aaggccaaag aagtttgccc catgtacttt atgaagctcc gctctggcct tatgataaaa 241 aaggaggcct gttactttag gagagaaacc accaaaaggc cttcactgaa aacaggaatt 301 tcacctatta cagagtatct tgcttctcta agcacataca atgatcaatc cattactttt 361 gctttggagg atgaaagtta tgagatatat gttgaagact tgaaaaaaga tgaaaagaaa 421 gataaggtgt tactgagtta ctatgagtct caacacccct caaatgaatc aggtgacggt 481 gttgatggta agatgttaat ggtaaccctg agtcctacaa aagacttctg gttgcatgcc 541 aacaacaagg aacactctgt ggagctccat aagtgtgaaa aaccactgcc agaccaggcc 601 ttctttgtcc ttcataatat gcactccaac tgtgtttcat ttgaatgcaa gactgatcct 661 ggagtgttta taggtgtaaa ggataatcat cttgctctga ttaaagtaga ctcttctgag 721 aatttgtgta ctgaaaatat cttgtttaag ctctctgaaa cttagttgat ggaaacctgt 781 gagtcttggg ttgagtaccc aaatgctacc actggagaag gaatgagaga taaagaaaga 841 gacaggtgac atctaaggga aatgaagagt gcttagcatg tgtggaatgt tttccatatt 901 atgtataaaa atattttttc taatcctcca gttattcttt tatttccctc tgtataactg 961 catcttcaat acaagtatca gtatattaaa tagggtattg gtaaagaaac ggtcaacatt 1021 ctaaagagat acagtctgac ctttactttt ctctagtttc agtccagaaa gaacttcata 1081 tttagagcta aggccactga ggaaagagcc atagcttaag tctctatgta gacagggatc 1141 cattttaaag agctacttag agaaataatt ttccacagtt ccaaacgata ggctcaaaca 1201 ctagagctgc tagtaaaaag aagaccagat gcttcacaga attatcattt tttcaactgg 1261 aataaaacac caggtttgtt tgtagatgtc ttaggcaaca ctcagagcag atctccctta 1321 ctgtcagggg atatggaact tcaaaggccc acatggcaag ccaggtaaca taaatgtgtg 1381 aaaaagtaaa gataactaaa aaatttagaa aaataaatcc agtatttgta aagtgaataa 1441 cttcatttct aattgtttaa tttttaaaat tctgattttt atatattgag tttaagcaag 1501 gcattcttac acgaggaagt gaagtaaatt ttagttcaga cataaaattt cacttattag 1561 gaatatgtaa catgctaaaa cttttttttt tttaaagagt actgagtcac aacatgtttt 1621 agagcatcca agtaccatat aatccaacta tcatggtaag gccagaaatc ttctaaccta 1681 ccagagccta gatgagacac cgaattaaca ttaaaatttc agtaactgac tgtccctcat 1741 gtccatggcc taccatccct tctgaccctg gcttccaggg acctatgtct tttaatactc 1801 actgtcacat tgggcaaagt tgcttctaat ccttatttcc catgtgcaca agtctttttg 1861 tattccagct tcctgataac actgcttact gtggaatatt catttgacat ctgtctcttt 1921 tcatttcttt taactaccat gcccttgata tatcttttgc acctgctgaa cttcatttct 1981 gtatcacctg acctctggat gccaaaacgt ttattctgct ttgtctgttg tagaatttta 2041 gataaagcta ttaatggcaa tatttttttg ctaaacgttt ttgtttttta ctgtcactag 2101 ggcaataaaa tttatactca accatataat aacatttttt aactactaaa ggagtagttt 2161 ttattttaaa gtcttagcaa tttctattac aacttttctt agacttaaca cttatgataa 2221 atgactaaca tagtaacaga atctttatga aatatgacct tttctgaaaa tacatacttt 2281 tacatttcta ctttattgag acctattaga tgtaagtgct agtagaatat aagataaaag 2341 aggctgagaa ttaccataca agggtattac aactgtaaaa caatttatct ttgtttcatt 2401 gttctgtcaa taattgttac caaagagata aaaataaaag cagaatgtat atcatcccat 2461 ctgaaaaaca ctaattattg acatgtgcat ctgtacaata aacttaaaat gattattaaa 2521 taatcaaata tatctactac attgtttata ttattgaata aagtatattt tccaaatgta 2581 aaaaaaaaaa aa Human IL-1R1 mRNA Variant 1 (SEQ ID NO: 104) 1 gtggccggcg gccggagccg actcggagcg cgcggcgccg gccgggagga gccggagagc 61 ggccgggccg ggcggtgggg gcgccggcct gccccgcgcg ccccagggag cggcaggaat 121 gtgacaatcg cgcgcccgcg caccgaagca ctcctcgctc ggctcctagg gctctcgccc 181 ctctgagctg agccgggttc cgcccggggc tgggatccca tcaccctcca cggccgtccg 241 tccaggtaga cgcaccctct gaagatggtg actccctcct gagaagctgg accccttggt 301 aaaagacaag gccttctcca agaagaatat gaaagtgtta ctcagactta tttgtttcat 361 agctctactg atttcttctc tggaggctga taaatgcaag gaacgtgaag aaaaaataat 421 tttagtgtca tctgcaaatg aaattgatgt tcgtccctgt cctcttaacc caaatgaaca 481 caaaggcact ataacttggt ataaagatga cagcaagaca cctgtatcta cagaacaagc 541 ctccaggatt catcaacaca aagagaaact ttggtttgtt cctgctaagg tggaggattc 601 aggacattac tattgcgtgg taagaaattc atcttactgc ctcagaatta aaataagtgc 661 aaaatttgtg gagaatgagc ctaacttatg ttataatgca caagccatat ttaagcagaa 721 actacccgtt gcaggagacg gaggacttgt gtgcccttat atggagtttt ttaaaaatga 781 aaataatgag ttacctaaat tacagtggta taaggattgc aaacctctac ttcttgacaa 841 tatacacttt agtggagtca aagataggct catcgtgatg aatgtggctg aaaagcatag 901 agggaactat acttgtcatg catcctacac atacttgggc aagcaatatc ctattacccg 961 ggtaatagaa tttattactc tagaggaaaa caaacccaca aggcctgtga ttgtgagccc 1021 agctaatgag acaatggaag tagacttggg atcccagata caattgatct gtaatgtcac 1081 cggccagttg agtgacattg cttactggaa gtggaatggg tcagtaattg atgaagatga 1141 cccagtgcta ggggaagact attacagtgt ggaaaatcct gcaaacaaaa gaaggagtac 1201 cctcatcaca gtgcttaata tatcggaaat tgaaagtaga ttttataaac atccatttac 1261 ctgttttgcc aagaatacac atggtataga tgcagcatat atccagttaa tatatccagt 1321 cactaatttc cagaagcaca tgattggtat atgtgtcacg ttgacagtca taattgtgtg 1381 ttctgttttc atctataaaa tcttcaagat tgacattgtg ctttggtaca gggattcctg 1441 ctatgatttt ctcccaataa aagcttcaga tggaaagacc tatgacgcat atatactgta 1501 tccaaagact gttggggaag ggtctacctc tgactgtgat atttttgtgt ttaaagtctt 1561 gcctgaggtc ttggaaaaac agtgtggata taagctgttc atttatggaa gggatgacta 1621 cgttggggaa gacattgttg aggtcattaa tgaaaacgta aagaaaagca gaagactgat 1681 tatcatttta gtcagagaaa catcaggctt cagctggctg ggtggttcat ctgaagagca 1741 aatagccatg tataatgctc ttgttcagga tggaattaaa gttgtcctgc ttgagctgga 1801 gaaaatccaa gactatgaga aaatgccaga atcgattaaa ttcattaagc agaaacatgg 1861 ggctatccgc tggtcagggg actttacaca gggaccacag tctgcaaaga caaggttctg 1921 gaagaatgtc aggtaccaca tgccagtcca gcgacggtca ccttcatcta aacaccagtt 1981 actgtcacca gccactaagg agaaactgca aagagaggct cacgtgcctc tcgggtagca 2041 tggagaagtt gccaagagtt ctttaggtgc ctcctgtctt atggcgttgc aggccaggtt 2101 atgcctcatg ctgacttgca gagttcatgg aatgtaacta tatcatcctt tatccctgag 2161 gtcacctgga atcagattat taagggaata agccatgacg tcaatagcag cccagggcac 2221 ttcagagtag agggcttggg aagatctttt aaaaaggcag taggcccggt gtggtggctc 2281 acgcctataa tcccagcact ttgggaggct gaagtgggtg gatcaccaga ggtcaggagt 2341 tcgagaccag cccagccaac atggcaaaac cccatctcta ctaaaaatac aaaaatgagc 2401 taggcatggt ggcacacgcc tgtaatccca gctacacctg aggctgaggc aggagaattg 2461 cttgaaccgg ggagacggag gttgcagtga gccgagtttg ggccactgca ctctagcctg 2521 gcaacagagc aagactccgt ctcaaaaaaa gggcaataaa tgccctctct gaatgtttga 2581 actgccaaga aaaggcatgg agacagcgaa ctagaagaaa gggcaagaag gaaatagcca 2641 ccgtctacag atggcttagt taagtcatcc acagcccaag ggcggggcta tgccttgtct 2701 ggggaccctg tagagtcact gaccctggag cggctctcct gagaggtgct gcaggcaaag 2761 tgagactgac acctcactga ggaagggaga catattcttg gagaactttc catctgcttg 2821 tattttccat acacatcccc agccagaagt tagtgtccga agaccgaatt ttattttaca 2881 gagcttgaaa actcacttca atgaacaaag ggattctcca ggattccaaa gttttgaagt 2941 catcttagct ttccacagga gggagagaac ttaaaaaagc aacagtagca gggaattgat 3001 ccacttctta atgctttcct ccctggcatg accatcctgt cctttgttat tatcctgcat 3061 tttacgtctt tggaggaaca gctccctagt ggcttcctcc gtctgcaatg tcccttgcac 3121 agcccacaca tgaaccatcc ttcccatgat gccgctcttc tgtcatcccg ctcctgctga 3181 aacacctccc aggggctcca cctgttcagg agctgaagcc catgctttcc caccagcatg 3241 tcactcccag accacctccc tgccctgtcc tccagcttcc cctcgctgtc ctgctgtgtg 3301 aattcccagg ttggcctggt ggccatgtcg cctgccccca gcactcctct gtctctgctc 3361 ttgcctgcac ccttcctcct cctttgccta ggaggccttc tcgcattttc tctagctgat 3421 cagaatttta ccaaaattca gaacatcctc caattccaca gtctctggga gactttccct 3481 aagaggcgac ttcctctcca gccttctctc tctggtcagg cccactgcag agatggtggt 3541 gagcacatct gggaggctgg tctccctcca gctggaattg ctgctctctg agggagaggc 3601 tgtggtggct gtctctgtcc ctcactgcct tccaggagca atttgcacat gtaacataga 3661 tttatgtaat gctttatgtt taaaaacatt ccccaattat cttatttaat ttttgcaatt 3721 attctaattt tatatataga gaaagtgacc tattttttaa aaaaatcaca ctctaagttc 3781 tattgaacct aggacttgag cctccatttc tggcttctag tctggtgttc tgagtacttg 3841 atttcaggtc aataacggtc ccccctcact ccacactggc acgtttgtga gaagaaatga 3901 cattttgcta ggaagtgacc gagtctagga atgcttttat tcaagacacc aaattccaaa 3961 cttctaaatg ttggaatttt caaaaattgt gtttagattt tatgaaaaac tcttctactt 4021 tcatctattc tttccctaga ggcaaacatt tcttaaaatg tttcattttc attaaaaatg 4081 aaagccaaat ttatatgcca ccgattgcag gacacaagca cagttttaag agttgtatga 4141 acatggagag gacttttggt ttttatattt ctcgtattta atatgggtga acaccaactt 4201 ttatttggaa taataatttt cctcctaaac aaaaacacat tgagtttaag tctctgactc 4261 ttgcctttcc acctgctttc tcctgggccc gctttgcctg cttgaaggaa cagtgctgtt 4321 ctggagctgc tgttccaaca gacagggcct agctttcatt tgacacacag actacagcca 4381 gaagcccatg gagcagggat gtcacgtctt gaaaagccta ttagatgttt tacaaattta 4441 attttgcaga ttattttagt ctgtcatcca gaaaatgtgt cagcatgcat agtgctaaga 4501 aagcaagcca atttggaaac ttaggttagt gacaaaattg gccagagagt gggggtgatg 4561 atgaccaaga attacaagta gaatggcagc tggaatttaa ggagggacaa gaatcaatgg 4621 ataagcgtgg gtggaggaag atccaaacag aaaagtgcaa agttattccc catcttccaa 4681 gggttgaatt ctggaggaag aagacacatt cctagttccc cgtgaacttc ctttgactta 4741 ttgtccccac taaaacaaaa caaaaaactt ttaatgcctt ccacattaat tagattttct 4801 tgcagttttt ttatggcatt tttttaaaga tgccctaagt gttgaagaag agtttgcaaa 4861 tgcaacaaaa tatttaatta ccggttgtta aaactggttt agcacaattt atattttccc 4921 tctcttgcct ttcttatttg caataaaagg tattgagcca ttttttaaat gacatttttg 4981 ataaattatg tttgtactag ttgatgaagg agtttttttt aacctgttta tataattttg 5041 cagcagaagc caaatttttt gtatattaaa gcaccaaatt catgtacagc atgcatcacg 5101 gatcaataga ctgtacttat tttccaataa aattttcaaa ctttgtactg ttaaaaaaaa 5161 aaaaaaaaaa Human IL-1R1 mRNA Variant 2 (SEQ ID NO: 105) 1 attggcagct cttcacttgt atcttttcat atcaaaaatg ggaggtgaca cccagtttaa 61 ggaaaattcc aaggcatttg tctcgactaa tgtgaaagat gattacagtg gccagaggac 121 tgccaaggct ccttctcaag ctgcttgagt caatgagggt agacgcaccc tctgaagatg 181 gtgactccct cctgagaagc tggacccctt ggtaaaagac aaggccttct ccaagaagaa 241 tatgaaagtg ttactcagac ttatttgttt catagctcta ctgatttctt ctctggaggc 301 tgataaatgc aaggaacgtg aagaaaaaat aattttagtg tcatctgcaa atgaaattga 361 tgttcgtccc tgtcctctta acccaaatga acacaaaggc actataactt ggtataaaga 421 tgacagcaag acacctgtat ctacagaaca agcctccagg attcatcaac acaaagagaa 481 actttggttt gttcctgcta aggtggagga ttcaggacat tactattgcg tggtaagaaa 541 ttcatcttac tgcctcagaa ttaaaataag tgcaaaattt gtggagaatg agcctaactt 601 atgttataat gcacaagcca tatttaagca gaaactaccc gttgcaggag acggaggact 661 tgtgtgccct tatatggagt tttttaaaaa tgaaaataat gagttaccta aattacagtg 721 gtataaggat tgcaaacctc tacttcttga caatatacac tttagtggag tcaaagatag 781 gctcatcgtg atgaatgtgg ctgaaaagca tagagggaac tatacttgtc atgcatccta 841 cacatacttg ggcaagcaat atcctattac ccgggtaata gaatttatta ctctagagga 901 aaacaaaccc acaaggcctg tgattgtgag cccagctaat gagacaatgg aagtagactt 961 gggatcccag atacaattga tctgtaatgt caccggccag ttgagtgaca ttgcttactg 1021 gaagtggaat gggtcagtaa ttgatgaaga tgacccagtg ctaggggaag actattacag 1081 tgtggaaaat cctgcaaaca aaagaaggag taccctcatc acagtgctta atatatcgga 1141 aattgaaagt agattttata aacatccatt tacctgtttt gccaagaata cacatggtat 1201 agatgcagca tatatccagt taatatatcc agtcactaat ttccagaagc acatgattgg 1261 tatatgtgtc acgttgacag tcataattgt gtgttctgtt ttcatctata aaatcttcaa 1321 gattgacatt gtgctttggt acagggattc ctgctatgat tttctcccaa taaaagtctt 1381 gcctgaggtc ttggaaaaac agtgtggata taagctgttc atttatggaa gggatgacta 1441 cgttggggaa gacattgttg aggtcattaa tgaaaacgta aagaaaagca gaagactgat 1501 tatcatttta gtcagagaaa catcaggctt cagctggctg ggtggttcat ctgaagagca 1561 aatagccatg tataatgctc ttgttcagga tggaattaaa gttgtcctgc ttgagctgga 1621 gaaaatccaa gactatgaga aaatgccaga atcgattaaa ttcattaagc agaaacatgg 1681 ggctatccgc tggtcagggg actttacaca gggaccacag tctgcaaaga caaggttctg 1741 gaagaatgtc aggtaccaca tgccagtcca gcgacggtca ccttcatcta aacaccagtt 1801 actgtcacca gccactaagg agaaactgca aagagaggct cacgtgcctc tcgggtagca 1861 tggagaagtt gccaagagtt ctttaggtgc ctcctgtctt atggcgttgc aggccaggtt 1921 atgcctcatg ctgacttgca gagttcatgg aatgtaacta tatcatcctt tatccctgag 1981 gtcacctgga atcagattat taagggaata agccatgacg tcaatagcag cccagggcac 2041 ttcagagtag agggcttggg aagatctttt aaaaaggcag taggcccggt gtggtggctc 2101 acgcctataa tcccagcact ttgggaggct gaagtgggtg gatcaccaga ggtcaggagt 2161 tcgagaccag cccagccaac atggcaaaac cccatctcta ctaaaaatac aaaaatgagc 2221 taggcatggt ggcacacgcc tgtaatccca gctacacctg aggctgaggc aggagaattg 2281 cttgaaccgg ggagacggag gttgcagtga gccgagtttg ggccactgca ctctagcctg 2341 gcaacagagc aagactccgt ctcaaaaaaa gggcaataaa tgccctctct gaatgtttga 2401 actgccaaga aaaggcatgg agacagcgaa ctagaagaaa gggcaagaag gaaatagcca 2461 ccgtctacag atggcttagt taagtcatcc acagcccaag ggcggggcta tgccttgtct 2521 ggggaccctg tagagtcact gaccctggag cggctctcct gagaggtgct gcaggcaaag 2581 tgagactgac acctcactga ggaagggaga catattcttg gagaactttc catctgcttg 2641 tattttccat acacatcccc agccagaagt tagtgtccga agaccgaatt ttattttaca 2701 gagcttgaaa actcacttca atgaacaaag ggattctcca ggattccaaa gttttgaagt 2761 catcttagct ttccacagga gggagagaac ttaaaaaagc aacagtagca gggaattgat 2821 ccacttctta atgctttcct ccctggcatg accatcctgt cctttgttat tatcctgcat 2881 tttacgtctt tggaggaaca gctccctagt ggcttcctcc gtctgcaatg tcccttgcac 2941 agcccacaca tgaaccatcc ttcccatgat gccgctcttc tgtcatcccg ctcctgctga 3001 aacacctccc aggggctcca cctgttcagg agctgaagcc catgctttcc caccagcatg 3061 tcactcccag accacctccc tgccctgtcc tccagcttcc cctcgctgtc ctgctgtgtg 3121 aattcccagg ttggcctggt ggccatgtcg cctgccccca gcactcctct gtctctgctc 3181 ttgcctgcac ccttcctcct cctttgccta ggaggccttc tcgcattttc tctagctgat 3241 cagaatttta ccaaaattca gaacatcctc caattccaca gtctctggga gactttccct 3301 aagaggcgac ttcctctcca gccttctctc tctggtcagg cccactgcag agatggtggt 3361 gagcacatct gggaggctgg tctccctcca gctggaattg ctgctctctg agggagaggc 3421 tgtggtggct gtctctgtcc ctcactgcct tccaggagca atttgcacat gtaacataga 3481 tttatgtaat gctttatgtt taaaaacatt ccccaattat cttatttaat ttttgcaatt 3541 attctaattt tatatataga gaaagtgacc tattttttaa aaaaatcaca ctctaagttc 3601 tattgaacct aggacttgag cctccatttc tggcttctag tctggtgttc tgagtacttg 3661 atttcaggtc aataacggtc ccccctcact ccacactggc acgtttgtga gaagaaatga 3721 cattttgcta ggaagtgacc gagtctagga atgcttttat tcaagacacc aaattccaaa 3781 cttctaaatg ttggaatttt caaaaattgt gtttagattt tatgaaaaac tcttctactt 3841 tcatctattc tttccctaga ggcaaacatt tcttaaaatg tttcattttc attaaaaatg 3901 aaagccaaat ttatatgcca ccgattgcag gacacaagca cagttttaag agttgtatga 3961 acatggagag gacttttggt ttttatattt ctcgtattta atatgggtga acaccaactt 4021 ttatttggaa taataatttt cctcctaaac aaaaacacat tgagtttaag tctctgactc 4081 ttgcctttcc acctgctttc tcctgggccc gctttgcctg cttgaaggaa cagtgctgtt 4141 ctggagctgc tgttccaaca gacagggcct agctttcatt tgacacacag actacagcca 4201 gaagcccatg gagcagggat gtcacgtctt gaaaagccta ttagatgttt tacaaattta 4261 attttgcaga ttattttagt ctgtcatcca gaaaatgtgt cagcatgcat agtgctaaga 4321 aagcaagcca atttggaaac ttaggttagt gacaaaattg gccagagagt gggggtgatg 4381 atgaccaaga attacaagta gaatggcagc tggaatttaa ggagggacaa gaatcaatgg 4441 ataagcgtgg gtggaggaag atccaaacag aaaagtgcaa agttattccc catcttccaa 4501 gggttgaatt ctggaggaag aagacacatt cctagttccc cgtgaacttc ctttgactta 4561 ttgtccccac taaaacaaaa caaaaaactt ttaatgcctt ccacattaat tagattttct 4621 tgcagttttt ttatggcatt tttttaaaga tgccctaagt gttgaagaag agtttgcaaa 4681 tgcaacaaaa tatttaatta ccggttgtta aaactggttt agcacaattt atattttccc 4741 tctcttgcct ttcttatttg caataaaagg tattgagcca ttttttaaat gacatttttg 4801 ataaattatg tttgtactag ttgatgaagg agtttttttt aacctgttta tataattttg 4861 cagcagaagc caaatttttt gtatattaaa gcaccaaatt catgtacagc atgcatcacg 4921 gatcaataga ctgtacttat tttccaataa aattttcaaa ctttgtactg ttaaaaaaaa 4981 aaaaaaaaaa Human IL-1R1 mRNA Variant 3 (SEQ ID NO: 106) 1 attggcagct cttcacttgt atcttttcat atcaaaaatg ggaggtgaca cccagtttaa 61 ggaaaattcc aaggcatttg tctcgactaa tgtgaaagat gattacagtg gccagaggac 121 tgccaaggct ccttctcaag ctgcttgagt caatgagggt agacgcaccc tctgaagatg 181 gtgactccct cctgagaagc tggacccctt ggtaaaagac aaggccttct ccaagaagaa 241 tatgaaagtg ttactcagac ttatttgttt catagctcta ctgatttctt ctctggaggc 301 tgataaatgc aaggaacgtg aagaaaaaat aattttagtg tcatctgcaa atgaaattga 361 tgttcgtccc tgtcctctta acccaaatga acacaaaggc actataactt ggtataaaga 421 tgacagcaag acacctgtat ctacagaaca agcctccagg attcatcaac acaaagagaa 481 actttggttt gttcctgcta aggtggagga ttcaggacat tactattgcg tggtaagaaa 541 ttcatcttac tgcctcagaa ttaaaataag tgcaaaattt gtggagaatg agcctaactt 601 atgttataat gcacaagcca tatttaagca gaaactaccc gttgcaggag acggaggact 661 tgtgtgccct tatatggagt tttttaaaaa tgaaaataat gagttaccta aattacagtg 721 gtataaggat tgcaaacctc tacttcttga caatatacac tttagtggag tcaaagatag 781 gctcatcgtg atgaatgtgg ctgaaaagca tagagggaac tatacttgtc atgcatccta 841 cacatacttg ggcaagcaat atcctattac ccgggtaata gaatttatta ctctagagga 901 aaacaaaccc acaaggcctg tgattgtgag cccagctaat gagacaatgg aagtagactt 961 gggatcccag atacaattga tctgtaatgt caccggccag ttgagtgaca ttgcttactg 1021 gaagtggaat gggtcagtaa ttgatgaaga tgacccagtg ctaggggaag actattacag 1081 tgtggaaaat cctgcaaaca aaagaaggag taccctcatc acagtgctta atatatcgga 1141 aattgaaagt agattttata aacatccatt tacctgtttt gccaagaata cacatggtat 1201 agatgcagca tatatccagt taatatatcc agtcactaat ttccagaagc acatgattgg 1261 tatatgtgtc acgttgacag tcataattgt gtgttctgtt ttcatctata aaatcttcaa 1321 gattgacatt gtgctttggt acagggattc ctgctatgat tttctcccaa taaaagcttc 1381 agatggaaag acctatgacg catatatact gtatccaaag actgttgggg aagggtctac 1441 ctctgactgt gatatttttg tgtttaaagt cttgcctgag gtcttggaaa aacagtgtgg 1501 atataagctg ttcatttatg gaagggatga ctacgttggg gaagacattg ttgaggtcat 1561 taatgaaaac gtaaagaaaa gcagaagact gattatcatt ttagtcagag aaacatcagg 1621 cttcagctgg ctgggtggtt catctgaaga gcaaatagcc atgtataatg ctcttgttca 1681 ggatggaatt aaagttgtcc tgcttgagct ggagaaaatc caagactatg agaaaatgcc 1741 agaatcgatt aaattcatta agcagaaaca tggggctatc cgctggtcag gggactttac 1801 acagggacca cagtctgcaa agacaaggtt ctggaagaat gtcaggtacc acatgccagt 1861 ccagcgacgg tcaccttcat ctaaacacca gttactgtca ccagccacta aggagaaact 1921 gcaaagagag gctcacgtgc ctctcgggta gcatggagaa gttgccaaga gttctttagg 1981 tgcctcctgt cttatggcgt tgcaggccag gttatgcctc atgctgactt gcagagttca 2041 tggaatgtaa ctatatcatc ctttatccct gaggtcacct ggaatcagat tattaaggga 2101 ataagccatg acgtcaatag cagcccaggg cacttcagag tagagggctt gggaagatct 2161 tttaaaaagg cagtaggccc ggtgtggtgg ctcacgccta taatcccagc actttgggag 2221 gctgaagtgg gtggatcacc agaggtcagg agttcgagac cagcccagcc aacatggcaa 2281 aaccccatct ctactaaaaa tacaaaaatg agctaggcat ggtggcacac gcctgtaatc 2341 ccagctacac ctgaggctga ggcaggagaa ttgcttgaac cggggagacg gaggttgcag 2401 tgagccgagt ttgggccact gcactctagc ctggcaacag agcaagactc cgtctcaaaa 2461 aaagggcaat aaatgccctc tctgaatgtt tgaactgcca agaaaaggca tggagacagc 2521 gaactagaag aaagggcaag aaggaaatag ccaccgtcta cagatggctt agttaagtca 2581 tccacagccc aagggcgggg ctatgccttg tctggggacc ctgtagagtc actgaccctg 2641 gagcggctct cctgagaggt gctgcaggca aagtgagact gacacctcac tgaggaaggg 2701 agacatattc ttggagaact ttccatctgc ttgtattttc catacacatc cccagccaga 2761 agttagtgtc cgaagaccga attttatttt acagagcttg aaaactcact tcaatgaaca 2821 aagggattct ccaggattcc aaagttttga agtcatctta gctttccaca ggagggagag 2881 aacttaaaaa agcaacagta gcagggaatt gatccacttc ttaatgcttt cctccctggc 2941 atgaccatcc tgtcctttgt tattatcctg cattttacgt ctttggagga acagctccct 3001 agtggcttcc tccgtctgca atgtcccttg cacagcccac acatgaacca tccttcccat 3061 gatgccgctc ttctgtcatc ccgctcctgc tgaaacacct cccaggggct ccacctgttc 3121 aggagctgaa gcccatgctt tcccaccagc atgtcactcc cagaccacct ccctgccctg 3181 tcctccagct tcccctcgct gtcctgctgt gtgaattccc aggttggcct ggtggccatg 3241 tcgcctgccc ccagcactcc tctgtctctg ctcttgcctg cacccttcct cctcctttgc 3301 ctaggaggcc ttctcgcatt ttctctagct gatcagaatt ttaccaaaat tcagaacatc 3361 ctccaattcc acagtctctg ggagactttc cctaagaggc gacttcctct ccagccttct 3421 ctctctggtc aggcccactg cagagatggt ggtgagcaca tctgggaggc tggtctccct 3481 ccagctggaa ttgctgctct ctgagggaga ggctgtggtg gctgtctctg tccctcactg 3541 ccttccagga gcaatttgca catgtaacat agatttatgt aatgctttat gtttaaaaac 3601 attccccaat tatcttattt aatttttgca attattctaa ttttatatat agagaaagtg 3661 acctattttt taaaaaaatc acactctaag ttctattgaa cctaggactt gagcctccat 3721 ttctggcttc tagtctggtg ttctgagtac ttgatttcag gtcaataacg gtcccccctc 3781 actccacact ggcacgtttg tgagaagaaa tgacattttg ctaggaagtg accgagtcta 3841 ggaatgcttt tattcaagac accaaattcc aaacttctaa atgttggaat tttcaaaaat 3901 tgtgtttaga ttttatgaaa aactcttcta ctttcatcta ttctttccct agaggcaaac 3961 atttcttaaa atgtttcatt ttcattaaaa atgaaagcca aatttatatg ccaccgattg 4021 caggacacaa gcacagtttt aagagttgta tgaacatgga gaggactttt ggtttttata 4081 tttctcgtat ttaatatggg tgaacaccaa cttttatttg gaataataat tttcctccta 4141 aacaaaaaca cattgagttt aagtctctga ctcttgcctt tccacctgct ttctcctggg 4201 cccgctttgc ctgcttgaag gaacagtgct gttctggagc tgctgttcca acagacaggg 4261 cctagctttc atttgacaca cagactacag ccagaagccc atggagcagg gatgtcacgt 4321 cttgaaaagc ctattagatg ttttacaaat ttaattttgc agattatttt agtctgtcat 4381 ccagaaaatg tgtcagcatg catagtgcta agaaagcaag ccaatttgga aacttaggtt 4441 agtgacaaaa ttggccagag agtgggggtg atgatgacca agaattacaa gtagaatggc 4501 agctggaatt taaggaggga caagaatcaa tggataagcg tgggtggagg aagatccaaa 4561 cagaaaagtg caaagttatt ccccatcttc caagggttga attctggagg aagaagacac 4621 attcctagtt ccccgtgaac ttcctttgac ttattgtccc cactaaaaca aaacaaaaaa 4681 cttttaatgc cttccacatt aattagattt tcttgcagtt tttttatggc atttttttaa 4741 agatgcccta agtgttgaag aagagtttgc aaatgcaaca aaatatttaa ttaccggttg 4801 ttaaaactgg tttagcacaa tttatatttt ccctctcttg cctttcttat ttgcaataaa 4861 aggtattgag ccatttttta aatgacattt ttgataaatt atgtttgtac tagttgatga 4921 aggagttttt tttaacctgt ttatataatt ttgcagcaga agccaaattt tttgtatatt 4981 aaagcaccaa attcatgtac agcatgcatc acggatcaat agactgtact tattttccaa 5041 taaaattttc aaactttgta ctgttaaaaa aaaaaaaaaa aaa Human IL-1R1 mRNA Variant 4 (SEQ ID NO: 107) 1 attaaagccc taagaggctg tgacacagcc atctccaaaa ccccactttc tccttccttt 61 gagcctccgt accagctggg gcgtccggca agatgtgagt tgtcactctg ctgcggcaca 121 gacctgaatt aacaactcta gctagggctg acttcaaaaa gcactttcgt tttttaataa 181 ccaacatcag ctcagcaggc ttcatttggg aaaagaaacc ttgtcggatt accccgacat 241 tctccacctc ctgggaggcc agccattccc aaatgcccca aggatgaaga acggagacgg 301 tagacgcacc ctctgaagat ggtgactccc tcctgagaag ctggacccct tggtaaaaga 361 caaggccttc tccaagaaga atatgaaagt gttactcaga cttatttgtt tcatagctct 421 actgatttct tctctggagg ctgataaatg caaggaacgt gaagaaaaaa taattttagt 481 gtcatctgca aatgaaattg atgttcgtcc ctgtcctctt aacccaaatg aacacaaagg 541 cactataact tggtataaag atgacagcaa gacacctgta tctacagaac aagcctccag 601 gattcatcaa cacaaagaga aactttggtt tgttcctgct aaggtggagg attcaggaca 661 ttactattgc gtggtaagaa attcatctta ctgcctcaga attaaaataa gtgcaaaatt 721 tgtggagaat gagcctaact tatgttataa tgcacaagcc atatttaagc agaaactacc 781 cgttgcagga gacggaggac ttgtgtgccc ttatatggag ttttttaaaa atgaaaataa 841 tgagttacct aaattacagt ggtataagga ttgcaaacct ctacttcttg acaatataca 901 ctttagtgga gtcaaagata ggctcatcgt gatgaatgtg gctgaaaagc atagagggaa 961 ctatacttgt catgcatcct acacatactt gggcaagcaa tatcctatta cccgggtaat 1021 agaatttatt actctagagg aaaacaaacc cacaaggcct gtgattgtga gcccagctaa 1081 tgagacaatg gaagtagact tgggatccca gatacaattg atctgtaatg tcaccggcca 1141 gttgagtgac attgcttact ggaagtggaa tgggtcagta attgatgaag atgacccagt 1201 gctaggggaa gactattaca gtgtggaaaa tcctgcaaac aaaagaagga gtaccctcat 1261 cacagtgctt aatatatcgg aaattgaaag tagattttat aaacatccat ttacctgttt 1321 tgccaagaat acacatggta tagatgcagc atatatccag ttaatatatc cagtcactaa 1381 tttccagaag cacatgattg gtatatgtgt cacgttgaca gtcataattg tgtgttctgt 1441 tttcatctat aaaatcttca agattgacat tgtgctttgg tacagggatt cctgctatga 1501 ttttctccca ataaaagctt cagatggaaa gacctatgac gcatatatac tgtatccaaa 1561 gactgttggg gaagggtcta cctctgactg tgatattttt gtgtttaaag tcttgcctga 1621 ggtcttggaa aaacagtgtg gatataagct gttcatttat ggaagggatg actacgttgg 1681 ggaagacatt gttgaggtca ttaatgaaaa cgtaaagaaa agcagaagac tgattatcat 1741 tttagtcaga gaaacatcag gcttcagctg gctgggtggt tcatctgaag agcaaatagc 1801 catgtataat gctcttgttc aggatggaat taaagttgtc ctgcttgagc tggagaaaat 1861 ccaagactat gagaaaatgc cagaatcgat taaattcatt aagcagaaac atggggctat 1921 ccgctggtca ggggacttta cacagggacc acagtctgca aagacaaggt tctggaagaa 1981 tgtcaggtac cacatgccag tccagcgacg gtcaccttca tctaaacacc agttactgtc 2041 accagccact aaggagaaac tgcaaagaga ggctcacgtg cctctcgggt agcatggaga DEMANDE OU BREVET VOLUMINEUX
LA PRESENTE PARTIE DE CETTE DEMANDE OU CE BREVET COMPREND
PLUS D'UN TOME.

NOTE : Pour les tomes additionels, veuillez contacter le Bureau canadien des brevets JUMBO APPLICATIONS/PATENTS
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Claims

CLAIMS:

1. A method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm in a subject, comprising:
releasing an immune modulator at a location in the gastrointestinal tract of the subject, wherein the method comprises administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of the immune modulator.

2. The method of claim 1, wherein the pharmaceutical composition is an ingestible device and the method comprises administering orally to the subject the pharmaceutical composition.

3. The method of claim 1 or 2, wherein the method does not comprise releasing more than 10% of the immune modulator at a location that is not proximate to the intended site of release.

4. The method of claim 1 or 2, wherein the method provides a concentration of the immune modulator at a location that is an intended site of release that is 2-100 times greater than at a location that is not the intended site of release.

5. The method of any one of the preceding claims, wherein the method provides a concentration of the immune modulator in the plasma of the subject that is less than 3 µg/mL.

6. The method of claim 5, wherein the method provides a concentration of the immune modulator in the plasma of the subject that is less than 0.3 µg/ml.

7. The method of claim 6, wherein the method provides a concentration of the immune modulator in the plasma of the subject that is less than 0.01 µg/mL.

8. The method of any one of claims 1 to 4, wherein the method provides a value of the immune modulator in the plasma of the subject that is less than 3 µg/ml.

9. The method of claim 8, wherein the method provides a C24 value of the immune modulator in the plasma of the subject that is less than 0.3 µg/mL.

10. The method of claim 9, wherein the method provides a C24 value of the immune modulator in the plasma of the subject that is less than 0.01 µg/mL.

11. The method of any one of claims 1 to 10, wherein the immune modulator is an inhibitory nucleic acid.

12. The method of claim 1 or 10, wherein the immune modulator is a small molecule.

13. The method of any one of claims 1 to 10, wherein the immune modulator is an antisense nucleic acid.

14. The method of any one of claims 1 to 10, wherein the immune modulator is a ribozyme.

15. The method of any one of claims 1 to 10, wherein the immune modulator is a siRNA.

16. The method of any one of claims 2 to 15, wherein the immune modulator is present in a pharmaceutical formulation within the device.

17. The method of claim 16, wherein the formulation is a solution of the immune modulator in a liquid medium.

18. The method of claim 17, wherein the formulation is a suspension of the immune modulator in a liquid medium.

19. The method of any one of claims 1 to 18, wherein the tissue originating from the endoderm is selected from the group consisting of: the stomach, the colon, the liver, the pancreas, the urinary bladder, the epithelial parts of the trachea, the lungs, the pharynx, the thyroid, the parathyroid, the intestines, and the gallbladder.

20. The method of any one of claims 1 to 18, wherein the inflammatory disease or condition originating from the endoderm is selected from the group consisting of: gastritis, Celiac disease, hepatitis, alcoholic lever disease, fatty liver disease (hepatic steatosis), non-alcoholic fatty liver disease (NASH), cirrhosis, primary schlerosing cholangitis, pancreatitis, insterstitial cystitits, asthma, chronic obstructic pulmonary disease, pulmonary fibrosis, pharyngitis, thyroiditis, hyperthyroidism, parathyroiditis, nephritis, Hashimoto's disease, Addison's disease, Graves' disease, Sjögren syndrome, type 1 diabetes, pelvic inflammatory disease, auditory canal inflammation, tinnitus, vestibular neuritis, otitis media, auditory canal inflammation, tracheitis, cholestatic liver disease, primary biliary schlerosis, liver parenchyma, an inherited metabolic disorder of the liver, Byler syndrome, cerebrotendinous, xanthomatosis, Zellweger's syndrome, neonatal hepatitis, cystic fibrosis, ALGS
(Alagilles syndrome), PFIC (progressive familial intrahepatic cholestasis), autoimmune hepatitis, primary biliary cirrhosis (PBC), liver fibrosis, NAFLD, portal hypertension, general cholestasis, such as in jaundice due to drugs or during pregnancy, intra- and extrahepatic cholestasis, such as hereditary forms of cholestasis, such as PFIC1, gall stones and choledocholithiasis, malignancy causing obstruction of the biliary tree, symptoms (scratching, pruritus) due to cholestasis/jaundice, chronic autoimmune liver disease leading to progressive cholestasis, and pruritus of cholestatic liver disease, duodenal ulcers, enteritis (radition-, chemotherapy-, or infection-induced enteritis), diverticulitis, pouchitis, cholecystitis, and cholangitis.

21. The method of any one of claims 1 to 19, wherein the inflammatory disease or condition that arises in a tissue originating from the endoderm is inflammation of the liver.

22. The method of any one of claims 1 to 21, wherein the immune modulator is released at a location in the large intestine of the subject.

23. The method of claim 22, wherein the location is in the proximal portion of the large intestine.

24. The method of claim 22, wherein the location is in the distal portion of the large intestine.

25. The method of any one of claims 1 to 21, wherein the immune modulator is released at a location in the ascending colon of the subject.

26. The method of claim 25, wherein the location is in the proximal portion of the ascending colon.

27. The method of claim 25, wherein the location is in the distal portion of the ascending colon.

28. The method of any one of claims 1 to 21, wherein the immune modulator is released at a location in the cecum of the subject.

29. The method of claim 28, wherein the location is in the proximal portion of the cecum.

30. The method of claim 28, wherein the location is in the distal portion of the cecum.

31. The method of any one of claims 1 to 21, wherein the immune modulator is released at a location in the sigmoid colon of the subject.

32. The method of claim 31, wherein the location is in the proximal portion of the sigmoid colon.

33. The method of claim 31, wherein the location is in the distal portion of the sigmoid colon.

34. The method of any one of claims 1 to 21, wherein the immune modulator is released at a location in the transverse colon of the subject.

35. The method of claim 34, wherein the location is in the proximal portion of the transverse colon.

36. The method of claim 34, wherein the location is in the distal portion of the transverse colon.

37. The method of any one of claims 1 to 21, wherein the immune modulator is released at a location in the descending colon of the subject.

38. The method of claim 37, wherein the location is in the proximal portion of the descending colon.

39. The method of claim 37, wherein the location is in the distal portion of the descending colon.

40. The method of any one of claims 1 to 21, wherein the immune modulator is released at a location in the small intestine of the subject.

41. The method of claim 40, wherein the location is in the proximal portion of the small intestine.

42. The method of claim 40, wherein the location is in the distal portion of the small intestine.

43. The method of any one of claims 1 to 21, wherein the immune modulator is released at a location in the duodenum of the subject.

44. The method of claim 43, wherein the location is in the proximal portion of the duodenum.

45. The method of claim 43, wherein the location is in the distal portion of the duodenum.

46. The method of any one of claims 1 to 21, wherein the immune modulator is released at a location in the jejunum of the subject.

47. The method of claim 46, wherein the location is in the proximal portion of the jejunum.

48. The method of claim 46, wherein the location is in the distal portion of the jejunum.

49. The method of any one of claims 1 to 21, wherein the immune modulator is released at a location in the ileum of the subject.

50. The method of claim 49, wherein the location is in the proximal portion of the ileum.

51. The method of claim 49, wherein the location is in the distal portion of the ileum.

52. The method of any one of the preceding claims, wherein the location at which the immune modulator is released is 10 cm or less from an intended site of release.

53. The method of any one of the preceding claims, wherein the location at which the immune modulator is released is 5 cm or less from an intended site of release.

54. The method of any one of the preceding claims, wherein the location at which the immune modulator is released is 2 cm or less from an intended site of release.

55. The method of any one of the preceding claims, wherein the immune modulator is released by mucosal contact.

56. The method of any one of the preceding claims, wherein the immune modulator is delivered to the location by a process that does not comprise systemic transport of the immune modulator.

57. The method of any one of the preceding claims, further comprising identifying an intended site of release of the immune modulator using a method that comprises imaging of the gastrointestinal tract.

58. The method of claim any one of the preceding claims, wherein the method comprises identifying an intended site of release of the immune modulator, prior to administering the pharmaceutical composition.

59. The method of claim 58, wherein the method comprises releasing the immune modulator substantially at the same time as identifying the intended site of release of the immune modulator.

60. The method of any one of the preceding claims, comprising (a) identifying a subject having an inflammatory disease or condition that arises in a tissue originating from the endoderm, and (b) evaluating the subject for suitability to treatment.

61. The method of any one of claims 1 or 3 to 15 or 17 to 60, wherein releasing the immune modulator is triggered by one or more of: a pH in the jejunum from 6.1 to 7.2, a pH in the mid small bowel from 7.0 to 7.8, a pH in the ileum from 7.0 to 8.0, a pH in the right colon from 5.7 to 7.0, a pH in the mid colon from 5.7 to 7.4, a pH in the left colon from 6.3 to 7.7, such as 7Ø

62. The method of any one of claims 1 to 60, wherein releasing the immune modulator is not dependent on the pH at or in the vicinity of the location.

63. The method of any one of claims 1 or 3 to 15 or 17 to 60, wherein releasing the immune modulator is triggered by degradation of a release component located in the device.

64. The method of any one of claims 1 to 60, wherein releasing the immune modulator is not triggered by degradation of a release component located in the device.

65. The method of any one of claims 1 to 60, wherein releasing the immune modulator is not dependent on enzymatic activity at or in the vicinity of the location.

66. The method of any one of claims 1 to 60, wherein releasing the immune modulator is not dependent on bacterial activity at or in the vicinity of the location.

67. The method of any one of claims 1 to 60, wherein the composition comprises a plurality of electrodes comprising a coating, and releasing the immune modulator is triggered by an electric signal by the electrodes resulting from the interaction of the coating with an intended site of release of the immune modulator.

68. The method of any one of claims 1 to 60, wherein releasing the immune modulator is triggered by a remote electromagnetic signal.

69. The method of any one of claims 1 to 60, wherein releasing the immune modulator is triggered by generation in the composition of a gas in an amount sufficient to expel the immune modulator.

70. The method of any one of claims 1 to 60, wherein releasing the immune modulator is triggered by an electromagnetic signal generated within the device according to a pre-determined drug release profile.

71. The method of any one of claims 2 to 60, wherein the ingestible device comprises an ingestible housing, wherein a reservoir storing the immune modulator is attached to the housing.

72. The method of claim 71, further comprising:
detecting when the ingestible housing is proximate to an intended site of release, wherein releasing the immune modulator comprises releasing the therapeutically effective amount of the immune modulator from the reservoir proximate the respective intended site of release in response to the detection.

73. The method of claim 72, wherein detecting comprises detecting via one or more sensors coupled to the ingestible housing.

74. The method of claim 73, wherein the one or more sensors comprise a plurality of coated electrodes and wherein detecting comprises receiving an electric signal by one or more of the coated electrodes responsive to the one or more electrode contacting the respective intended site of release.

75. The method of claim 72, wherein releasing comprises opening one or more valves in fluid communication with the reservoir.

76. The method of claim 0, wherein the one or more valves is communicably coupled to a processor positioned in the housing, the processor communicably coupled to one or more sensors configured to detect the intended site of release.

77. The method of claim 72, wherein releasing comprises pumping the therapeutically effective amount of the immune modulator from the reservoir via pump positioned in the ingestible housing.

78. The method of claim 0, wherein the pump is communicably coupled to a processor positioned in the housing, the processor communicably coupled to one or more sensors configured to detect an intended site of release of the immune modulator.

79. The method of claim 71, wherein the therapeutically effective amount of the immune modulator is stored in the reservoir at a reservoir pressure higher than a pressure in the gastrointestinal tract of the subject.

80. The method of claim 71, further comprising anchoring the ingestible housing at a location proximate to the intended site of release in response to the detection.

81. The method of claim 0, wherein anchoring the ingestible housing comprises one or more legs to extend from the ingestible housing.

82. The method of any one of the preceding claims, wherein the amount of the immune modulator that is administered is from about 1 mg to about 500 mg.

83. The method of any one of the preceding claims, wherein the immune modulator is an antibody or an antigen-binding antibody fragment.

84. The method of claim 83, wherein the antibody is a humanized antibody.

85. The method of any one of claims 1 to 84, wherein the amount of the immune modulator is less than an amount that is effective when the immune modulator is administered systemically.

86. The method of any one of the preceding claims, comprising administering (i) an amount of the immune modulator that is an induction dose.

87. The method of claim 86, further comprising (ii) administering an amount of the immune modulator that is a maintenance dose following the administration of the induction dose.

88. The method of claim 86 or 87, wherein the induction dose is administered once a day.

89. The method of claim 86 or 87, wherein the induction dose is administered once every three days.

90. The method of claim 86 or 87, wherein the induction dose is administered once a week.

91. The method of claim 87, wherein step (ii) is repeated one or more times.

92. The method of claim 87, wherein step (ii) is repeated once a day over a period of about 6-8 weeks.

93. The method of claim 87, wherein step (ii) is repeated once every three days over a period of about 6-8 weeks.

94. The method of claim 87, wherein step (ii) is repeated once a week over a period of about 6-8 weeks.

95. The method of claim 87, wherein the induction dose is equal to the maintenance dose.

96. The method of claim 87, wherein the induction dose is greater than the maintenance dose.

97. The method of claim 87, wherein the induction dose is 5 times greater than the maintenance dose.

98. The method of claim 87, wherein the induction dose is 2 times greater than the maintenance dose.

99. The method of any one of the preceding claims, wherein the method comprises releasing the immune modulator at the location in the gastrointestinal tract as a single bolus.

100. The method of any one of claims 1 to 98, wherein the method comprises releasing the immune modulator at the location in the gastrointestinal tract as more than one bolus.

101. The method of any one of claims 1 to 98, wherein the method comprises delivering the immune modulator at the location in the gastrointestinal tract in a continuous manner.

102. The method of claim 101, wherein the method comprises delivering the immune modulator at the location in the gastrointestinal tract over a time period of 20 or more minutes.

103. The method of any one of claims 1 to 102, wherein the method does not comprise delivering an immune modulator rectally to the subject.

104. The method of any one of claims 1 to 102, wherein the method does not comprise delivering an immune modulator via an enema to the subject.

105. The method of any one of claims 1 to 102, wherein the method does not comprise delivering an immune modulator via suppository to the subject.

106. The method of any one of claims 1 to 102, wherein the method does not comprise delivering an immune modulator via instillation to the rectum of the subject.

107. The method of any one of claims 1 to 102, wherein the method does not comprise surgical implantation.

108. The method of any one of the preceding claims, wherein the immune modulator is a IL-12/IL-23 inhibitor.

109. The method of any one of the preceding claims, wherein the immune modulator is a TNF.alpha. inhibitor.

110. The method of any one of the preceding claims, wherein the immune modulator is a IL-6 receptor inhibitor.

111. The method of any one of the preceding claims, wherein the immune modulator is a CD40/CD40L inhibitor.

112. The method of any one of the preceding claims, wherein the immune modulator is a IL-1 inhibitor.

113. The method of any one of claims 1 to 67 or 69 to 112, wherein the composition is an autonomous device.

114. The method of any one of claims 1 to 113, wherein the composition comprises a mechanism capable of releasing the immune modulator.

115. The method of any one of claims 1 to 114, wherein the composition comprises a tissue anchoring mechanism for anchoring the composition to the location.

116. The method of claim 115, wherein the tissue anchoring mechanism is capable of activation for anchoring to the location.

117. The method of claim 115 to 116, wherein the tissue anchoring mechanism comprises an osmotically-driven sucker.

118. The method of claim 115, 116, or 117, wherein the tissue anchoring mechanism comprises a connector operable to anchor the composition to the location.

119. The method of claim 118, wherein the connector is operable to anchor the composition to the location using an adhesive, negative pressure and/or fastener.

120. The method of claim 71, wherein the reservoir is an anchorable reservoir.

121. The method of any one of claims 1 to 60, wherein the pharmaceutical composition is an ingestible device, comprising:
a housing;
a reservoir located within the housing and containing the immune modulator, a mechanism for releasing the immune modulator from the reservoir;
and;
an exit valve configured to allow the immune modulator to be released out of the housing from the reservoir.

122. The method of claim 121, wherein the ingestible device further comprises:

an electronic component located within the housing; and a gas generating cell located within the housing and adjacent to the electronic component, wherein the electronic component is configured to activate the gas generating cell to generate gas.

123. The method of claim 121 or 122, wherein the ingestible device further comprises:
a safety device placed within or attached to the housing, wherein the safety device is configured to relieve an internal pressure within the housing when the internal pressure exceeds a threshold level.

124. The method of claim 1 to 60, wherein the pharmaceutical composition is an ingestible device, comprising:
a housing defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end;
an electronic component located within the housing;
a gas generating cell located within the housing and adjacent to the electronic component, wherein the electronic component is configured to activate the gas generating cell to generate gas;
a reservoir located within the housing, wherein the reservoir stores a dispensable substance and a first end of the reservoir is attached to the first end of the housing;
an exit valve located at the first end of the housing, wherein the exit valve is configured to allow the dispensable substance to be released out of the first end of the housing from the reservoir; and a safety device placed within or attached to the housing, wherein the safety device is configured to relieve an internal pressure within the housing when the internal pressure exceeds a threshold level.

125. The method of claim 1 to 60, wherein the pharmaceutical composition is an ingestible device, comprising:
a housing defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end;
an electronic component located within the housing, a gas generating cell located within the housing and adjacent to the electronic component, wherein the electronic component is configured to activate the gas generating cell to generate gas;
a reservoir located within the housing, wherein the reservoir stores a dispensable substance and a first end of the reservoir is attached to the first end of the housing;
an injection device located at the first end of the housing, wherein the jet injection device is configured to inject the dispensable substance out of the housing from the reservoir; and a safety device placed within or attached to the housing, wherein the safety device is configured to relieve an internal pressure within the housing.

126. The method of claim 1 to 60, wherein the pharmaceutical composition is an ingestible device, comprising:
a housing defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end;
an optical sensing unit located on a side of the housing, wherein the optical sensing unit is configured to detect a reflectance from an environment external to the housing;
an electronic component located within the housing;
a gas generating cell located within the housing and adjacent to the electronic component, wherein the electronic component is configured to activate the gas generating cell to generate gas in response to identifying a location of the ingestible device based on the reflectance;
a reservoir located within the housing, wherein the reservoir stores a dispensable substance and a first end of the reservoir is attached to the first end of the housing;
a membrane in contact with the gas generating cell and configured to move or deform into the reservoir by a pressure generated by the gas generating cell; and a dispensing outlet placed at the first end of the housing, wherein the dispensing outlet is configured to deliver the dispensable substance out of the housing from the reservoir.

127. The method of any one of claims 1 to 60, wherein the pharmaceutical composition is an ingestible device as disclosed in US Patent Application Ser.
No.
62/385,553, incorporated by reference herein in its entirety.

128. The method of any one of claims 1 to 60, wherein the pharmaceutical composition is an ingestible device comprising a localization mechanism as disclosed in international patent application PCT/US2015/052500, incorporated by reference herein in its entirety.

129. The method of any one of claims 1 to 60, wherein the pharmaceutical composition is not a dart-like dosage form.

130. A method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm of a subject, comprising:
releasing an immune modulator at a location in the large intestine of the subject, wherein the method comprises administering endoscopically to the subject a therapeutically effective amount of the immune modulator, wherein the method does not comprise releasing more than 20% of the immune modulator at a location that is not an intended site of release.

131. A method of treating a disease or condition that arises in a tissue originating from the endoderm in a subject, comprising:
releasing an immune modulator at a location in the proximal portion of the large intestine of the subject, wherein the method comprises administering endoscopically to the subject a pharmaceutical composition comprising a therapeutically effective amount of the immune modulator, wherein the pharmaceutical composition is an ingestible device.

132. The method of claim 130 or 131, wherein the method does not comprise releasing more than 20% of the immune modulator at a location that is not proximate to an intended site of release.

133. The method of claim 130, 131 or 132 wherein the method does not comprise releasing more than 10% of the immune modulator at a location that is not proximate to an intended site of release.

134. The method of any one of claims 130, 131 or 132, wherein the method provides a concentration of the immune modulator at a location that is an intended site of release that is 2-100 times greater than at a location that is not the intended site of release.

135. The method of any one of claims 130 to 134, wherein the method provides a concentration of the immune modulator in the plasma of the subject that is less than 3 µg/mL.

136. The method of claim 135, wherein the method provides a concentration of the immune modulator in the plasma of the subject that is less than 0.3 µg/mL.

137. The method of claim 136, wherein the method provides a concentration of the immune modulator in the plasma of the subject that is less than 0.01 µg/mL.

138. The method of any one of claims 130 to 134, wherein the method provides a C24 value of the immune modulator in the plasma of the subject that is less than 3 µg/mL.

139. The method of any one of claims 130 to 134, wherein the method provides a C24 value of the immune modulator in the plasma of the subject that is less than 0.3 µg/mL.

140. The method of any one of claims 130 to 134, wherein the method provides a C24 value of the immune modulator in the plasma of the subject that is less than 0.01 µg/mL.

141. The method of any one of claims 130 to 134, wherein the composition does not comprise an enteric coating.

142. The method of any one of claims 130 to 141, wherein the immune modulator is not a cyclic peptide.

143. The method of any one of claims 130 to 141, wherein the immune modulator is present in a pharmaceutical formulation within the device.

144. The method of claim 143, wherein the formulation is a solution of the immune modulator in a liquid medium.

145. The method of claim 143, wherein the formulation is a suspension of the immune modulator in a liquid medium.

146. The method of any one of claims 130 to 145, wherein the tissue originating from the endoderm is selected from the group consisting of: the stomach, the colon, the liver, the pancreas, the urinary bladder, the epithelial parts of the trachea, the lungs, the pharynx, the thyroid, the parathyroid, the intestines, and the gallbladder.

147. The method of any one of claims 130 to 145, wherein the inflammatory disease or condition that arises in a tissue originating from the endoderm is selected from the group consisting of: gastritis, Celiac disease, hepatitis, alcoholic lever disease, fatty liver disease (hepatic steatosis), non-alcoholic fatty liver disease (NASH), cirrhosis, primary schlerosing cholangitis, pancreatitis, insterstitial cystitits, asthma, chronic obstructic pulmonary disease, pulmonary fibrosis, pharyngitis, thyroiditis, hyperthyroidism, parathyroiditis, nephritis, Hashimoto's disease, Addison's disease, Graves' disease, Sjögren syndrome, type 1 diabetes, pelvic inflammatory disease, auditory canal inflammation, tinnitus, vestibular neuritis, otitis media, auditory canal inflammation, tracheitis, cholestatic liver disease, primary biliary schlerosis, liver parenchyma, an inherited metabolic disorder of the liver, Byler syndrome, cerebrotendinous, xanthomatosis, Zellweger's syndrome, neonatal hepatitis, cystic fibrosis, ALGS (Alagilles syndrome), PFIC (progressive familial intrahepatic cholestasis), autoimmune hepatitis, primary biliary cirrhosis (PBC), liver fibrosis, NAFLD, portal hypertension, general cholestasis, such as in jaundice due to drugs or during pregnancy, intra- and extrahepatic cholestasis, such as hereditary forms of cholestasis, such as PFIC1, gall stones and choledocholithiasis, malignancy causing obstruction of the biliary tree, symptoms (scratching, pruritus) due to cholestasis/jaundice, chronic autoimmune liver disease leading to progressive cholestasis, and pruritus of cholestatic liver disease, duodenal ulcers, enteritis (radition-, chemotherapy-, or infection-induced enteritis), diverticulitis, pouchitis, cholecystitis, and cholangitis.

148. The method of any one of claims 130 to 145, wherein the inflammatory disease or condition that arises in a tissue originating from the endoderm is inflammation of the liver.

149. The method of any one of claims 130 to 148, wherein the immune modulator is released at a location in the proximal portion of the ascending colon.

150. The method of any one of claims 130 to 148, wherein the immune modulator is released at a location in the proximal portion of the cecum.

151. The method of any one of claims 130 to 148, wherein the immune modulator is released at a location in the proximal portion of the sigmoid colon.

152. The method of any one of claims 130 to 148, wherein the immune modulator is released at a location in the proximal portion of the transverse colon.

153. The method of any one of claims 130 to 148, wherein the immune modulator is released at a location in the proximal portion of the descending colon.

154. The method of any one of claims 130 to 148, wherein the method comprises administering to the subject a reservoir comprising the therapeutically effective amount of the immune modulator, wherein the reservoir is connected to the endoscope.

155. The method of any one of the preceding claims, further comprising administering a second agent orally, intravenously or subcutaneously, wherein the second agent is the same immune modulator; a different immune modulator; or an agent having a different biological target from the immune modulator, wherein the second agent is an agent suitable for treating an inflammatory disease or condition that arises in a tissue originating from the endoderm.

156. The method of claim 155, wherein the immune modulator is administered prior to the second agent.

157. The method of claim 155, wherein the immune modulator is administered after the second agent.

158. The method of claim 155, wherein the immune modulator and the second agent are administered substantially at the same time.

159. The method of any one of claims 155, wherein the second agent is administered intravenously.

160. The method of any one of claims 155, wherein the second agent is administered subcutaneously.

161. The method of any one of claims 155 to 160, wherein the amount of the second agent is less than the amount of the second agent when the immune modulator and the second agent are both administered systemically.

162. The method of claim 161, wherein the second agent is another immune modulator.

163. The method of any one of claims 1 to 154, wherein the method does not comprise administering a second agent.

164. The method of any one of claims 119 to 163, wherein the method comprises identifying an intended site of release prior to endoscopic administration.

165. The method of any one of claims 119 to 164, wherein the method comprises identifying an intended site of release substantially at the same time as releasing the immune modulator.

166. The method of any one of the preceding claims, wherein the method comprising monitoring the progress of the disease.

167. The method of any one of claims 1 to 164, wherein the method does not comprise administering an immune modulator with a spray catheter.

168. The method of any one of claims 1 to 164, wherein the method comprises administering an immune modulator with a spray catheter.

169. A method of treating an inflammatory disease or condition arising in a tissue originating from the endoderm in a subject, comprising:
releasing an immune modulator at a location in the gastrointestinal tract of the subject that is proximate to an intended site of release, wherein the method comprises administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of the immune modulator the method comprising one or more of the following steps:
a) identifying a subject having a disease or condition that arises in a tissue originating from the endoderm;
b) determination of the severity of the disease;
c) determination of the location of the disease;
d) evaluating the subject for suitability to treatment;
e) administration of an induction dose of the immune modulator;
monitoring the progress of the disease; and/or optionally repeating steps e) and f) one or more times.

170. The method of claim 169, wherein the pharmaceutical composition is an ingestible device and the method comprises administering orally to the subject the pharmaceutical composition.

171. The method of claim 169 or 170, wherein the method comprises administering one or more maintenance doses following administration of the induction dose in step e).

172. The method of claim 171, wherein the induction dose is a dose of the immune modulator administered in an ingestible device.

173. The method of claim 171 or 172, wherein the maintenance dose is a dose of the immune modulator administered in an ingestible device as disclosed herein.

174. The method of claim 171 or 172, wherein the maintenance dose is a dose of the immune modulator delivered systemically.

175. The method of claim 171, wherein the induction dose is a dose of the immune modulator delivered systemically.

176. The method of claim 171 or 175, wherein the maintenance dose is a dose of the immune modulator administered in an ingestible device.

177. The method of claim 171, wherein the induction dose is a dose of a second agent as delivered systemically.

178. The method of claim 171 or 175, wherein the maintenance dose is a dose of the immune modulator administered in an ingestible device.

179. An immune modulator delivery apparatus comprising:
an ingestible housing comprising a reservoir having a pharmaceutical composition comprising a therapeutically effective amount of the immune modulator stored therein;
a detector coupled to the ingestible housing, the detector configured to detect when the ingestible housing is proximate to a respective intended site of release;
a valve system in fluid communication with the reservoir system; and a controller communicably coupled to the valve system and the detector, the controller configured to cause the valve system to open in response to the detector detecting that the ingestible housing is proximate to the respective intended site of release so as to release the therapeutically effective amount of the immune modulator at the respective intended site of release.

180. The immune modulator delivery apparatus according to claim 179, further comprising a pump positioned in the ingestible housing, the pump configured to pump the therapeutically effective amount of the immune modulator from the reservoir in response to activation of the pump by the controller responsive to detection by the detector of the ingestible housing being proximate to the intended site of release.

181. The immune modulator delivery apparatus according to claim 180, wherein the controller is configured to cause the pump to pump the therapeutically effective amount of the immune modulator from the reservoir according to the following protocol.

182. The immune modulator delivery apparatus according to claim 179, wherein the valve system comprises a dissolvable coating.

183. The anti-inflammtaory agent delivery apparatus according to claim 179, wherein the valve system comprises one or more doors configured for actuation by at least one of sliding, pivoting, and rotating.

184. The immune modulator delivery apparatus according to claim 179, wherein the valve system comprises an electrostatic shield.

185. The immune modulator delivery apparatus according to claim 179, wherein the reservoir comprises a pressurized cell.

186. The immune modulator delivery apparatus according to claim 179, further comprising at least one actuatable anchor configured to retain the ingestible housing at the respective intended site of release upon actuation.

187. The anti-inflammatory inhibitor delivery apparatus according to claim 179, herein the actuatable anchor is retractable.

188. A composition comprising a therapeutically effective amount of the immune modulator of any one of the preceding claims, wherein the composition is capable of releasing the immune modulator at a location in the gastrointestinal tract of the subject.

189. The composition of claim 188, wherein the composition comprises a tissue anchoring mechanism for anchoring the composition to the location.

190. The composition of claim 189, wherein the tissue anchoring mechanism is capable of anchoring for anchoring to the location.

191. The composition of claim 189 or 190, wherein the tissue anchoring mechanism comprises an osmotically-driven sucker.

192. The composition of claim 189, 190 or 191, wherein the tissue anchoring mechanism comprises a connector operable to anchor the composition to the location.

193. The composition of claim 192, wherein the connector is operable to anchor the composition to the location using an adhesive, negative pressure and/or fastener.

194. An immune modulator for use in a method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm in a subject, wherein the method comprises orally administering to the subject an ingestible device loaded with the immune modulator, wherein the immune modulator is released by the device at a location in the gastrointestinal tract of the subject that is proximate to an intended site of release of the immune modulator.

195. The immune modulator for use of claim 194, wherein the immune modulator is contained in a reservoir suitable for attachment to a device housing, and wherein the method comprises attaching the reservoir to the device housing to form the ingestible device, prior to orally administering the ingestible device to the subject.

196. An attachable reservoir containing an immune modulator for use in a method of treating an inflammatory disease or condition that arises in a tissue originating from the endoderm, wherein the method comprises attaching the reservoir to a device housing to form an ingestible device and orally administering the ingestible device to a subject, wherein the immune modulator is released by device at a location in the gastrointestinal tract of the subject that is proximate to the intended site of release.

197. A composition comprising or consisting of an ingestible device loaded with a therapeutically effective amount of an immune modulator, for use in a method of treatment, wherein the method comprises orally administering the composition to the subject, wherein the immune modulator is released by the device at a location in the gastrointestinal tract of the subject that is proximate to an intended site of release.

198. The immune modulator for use according to claim 194 or 195, the attachable reservoir compartment for use according to claim 196, or the composition for use according to claim 197, wherein the intended site of release has been pre-determined.

199. The immune modulator for use according to claim 194 or 195, the attachable reservoir compartment for use according to claim 196, or the composition for use according to claim 197, wherein the ingestible device further comprises an environmental sensor and the method further comprises using the environmental sensor to identify the location of the intended site of release.

200. The immune modulator for use, the attachable reservoir compartment for use the composition for use, according to claim 199, wherein the environmental sensor is an imaging sensor and the method further comprising imaging the gastrointestinal tract to identify the intended site of release.

201. The immune modulator for use, the attachable reservoir compartment for use, or the composition for use, according to claim 200, wherein the imaging detects an intended site of release.

202. The immune modulator for use, the attachable reservoir compartment for use or the composition for use, according to any one of claims 194 to 201, wherein the inflammatory disease or condition that arises in a tissue originating from the endoderm is selected from the group of: gastritis, Celiac disease, hepatitis, alcoholic lever disease, fatty liver disease (hepatic steatosis), non-alcoholic fatty liver disease (NASH), cirrhosis, primary schlerosing cholangitis, pancreatitis, insterstitial cystitits, asthma, chronic obstructic pulmonary disease, pulmonary fibrosis, pharyngitis, thyroiditis, hyperthyroidism, parathyroiditis, nephritis, Hashimoto's disease, Addison's disease, Graves' disease, Sjögren syndrome, type 1 diabetes, pelvic inflammatory disease, auditory canal inflammation, tinnitus, vestibular neuritis, otitis media, auditory canal inflammation, tracheitis, cholestatic liver disease, primary biliary schlerosis, liver parenchyma, an inherited metabolic disorder of the liver, Byler syndrome, cerebrotendinous, xanthomatosis, Zellweger's syndrome, neonatal hepatitis, cystic fibrosis, ALGS (Alagilles syndrome), PFIC (progressive familial intrahepatic cholestasis), autoimmune hepatitis, primary biliary cirrhosis (PBC), liver fibrosis, NAFLD, portal hypertension, general cholestasis, such as in jaundice due to drugs or during pregnancy, intra- and extrahepatic cholestasis, such as hereditary forms of cholestasis, such as PFIC1, gall stones and choledocholithiasis, malignancy causing obstruction of the biliary tree, symptoms (scratching, pruritus) due to cholestasis/jaundice, chronic autoimmune liver disease leading to progressive cholestasis, and pruritus of cholestatic liver disease, duodenal ulcers, enteritis (radition-, chemotherapy-, or infection-induced enteritis), diverticulitis, pouchitis, cholecystitis, and cholangitis.

203. An ingestible device loaded with a therapeutically effective amount of an immune modulator, wherein the device is controllable to release the immune modulator at a location in the gastrointestinal tract of the subject that is proximate to an intended site of release.

204. The device of claim 203 for use in a method of treatment of the human or animal body.

205. The immune modulator for use, the attachable reservoir compartment for use or the composition for use according to any one of claims 194 to 202, or the device according to claim 203 or claim 204, wherein the ingestible device comprises:
a housing defined by a first end, a second end substantially opposite from the first end, and a wall extending longitudinally from the first end to the second end;
a reservoir located within the housing and containing the immune modulator wherein a first end of the reservoir is connected to the first end of the housing;
a mechanism for releasing the anti-inflammatory from the reservoir; and an exit value configured to allow the immune modulator to be released out of the housing from the reservoir.

206. The immune modulator for use, the attachable reservoir compartment for use or the composition for use according to any one of claims 194 to 202, or the device according to claim 203 or claim 204, wherein the ingestible device comprises:
an ingestible housing comprising a reservoir compartment having a therapeutically effective amount of the immune modulator stored therein;
a release mechanism having a closed state which retains the immune modulator in the reservoir and an open state which releases the immune modulator the reservoir to the exterior of the device; and an actuator which changes the state of the release mechanism from the closed to the open state.

207. The immune modulator for use, the attachable reservoir compartment for use, the composition for use, or the device according to claims 205 or 206, wherein the ingestible device further comprises an environmental sensor for detecting the location of the device in the gut.

208. The immune modulator for use, the attachable reservoir compartment for use, the composition for use, or the device according to claim 207, wherein the ingestible device further comprises a communication system for transmitting data from the environmental sensor to an external receiver.

209. The immune modulator for use, the attachable reservoir compartment for use, the composition for use, or the device according to claim 207 or 208, wherein the ingestible device further comprises a processor or controller which is coupled to the environmental sensor and to the actuator and which triggers the actuator to cause the release mechanism to transition from its closed state to its open state when it is determined that the device is at or proximal to the intended site of release.

210. The immune modulator for use, the attachable reservoir compartment for use, the composition for use, or the device according to claim 208, wherein the communication system further comprises means for receiving a signal from an external transmitter, and wherein the actuator is adapted to be triggered in response to the signal.

211. The immune modulator for use, the attachable reservoir compartment for use, the composition for use, or the device according to any one of claims 205 to 210, wherein the ingestible device further comprises a communication system for transmitting localization data to an external receiver.

212. The immune modulator for use, the attachable reservoir compartment for use, the composition for use, or the device according to any one of claims 205 to 208, wherein the ingestible device further comprises a communication system for transmitting localization data to an external receiver and for receiving a signal from an external transmitter; wherein the actuator is adapted to be triggered in response to the signal.

213. The immune modulator for use, the attachable reservoir compartment for use, the composition for use, or the device according to any one of claims 114 to 212, wherein the ingestible device further comprises a deployable anchoring system and an actuator for deploying the anchoring system, wherein the anchoring system is capable of anchoring or attaching the ingestible device to the subject's tissue.

214. The method of any one of claims 1-178, wherein the subject has previously been identified as having an inflammatory disease or condition that arises in a tissue originating from the endoderm.

215. The method of any one of claims 1-107 or 113-178, wherein the immune modulator is selected from the group consisting of: IL-12/IL-23 inhibitors, TNF.alpha. inhibitors, IL-6 receptor inhibitors, CD40/CD40L inhibitors, IL-1 inhibitors, IL-13 inhibitors, IL-10 receptor agonists, and integrin inhibitors.

216. A method of formulating a pharmaceutical composition comprising an immune modulator, the method comprising:
(a) topically administering a dose of an immune modulator to a small intestine and/or colon of a mammal;
(b) selecting an immune modulator whose topical administration in step (a) has been determined to result in (i) a decrease in one or both of the level of T cells in a mesenteric lymph node and the level of T cells in a Peyer's patch in the mammal, and/or (ii) an increase in the level of T cells in blood in the mammal, each as compared to the corresponding level in a control mammal systemically administered the same dose of the immune modulator; and (c) formulating a pharmaceutical composition comprising the selected immune modulator.

217. A method of formulating a pharmaceutical composition comprising an immune modulator, the method comprising:
(a) selecting an immune modulator whose topical administration to a small intestine and/or colon of a mammal has been determined to result in (i) a decrease in one or both of the level of T cells in a mesenteric lymph node and the level of T cells in a Peyer's patch in the mammal, and/or (ii) an increase in the level of T cells in blood in the mammal, each as compared to the corresponding level in a control mammal systemically administered the same dose of the immune modulator; and (b) formulating a pharmaceutical composition comprising the selected immune modulator.

218. A method of formulating a pharmaceutical composition comprising an immune modulator, the method comprising formulating a pharmaceutical composition comprising an immune modulator determined to result in a mammal topically administered a dose of an immune modulator to the small intestine and/or colon of the mammal:
(i) a decrease in one or both of the level of T cells in a mesenteric lymph node and the level of T
cells in a Peyer's patch in a mammal, and/or (ii) an increase in the level of T cells in blood in the mammal, each as compared to the corresponding level in a control mammal systemically administered the same dose of the immune modulator.

219. The method of any one of claim 216-218, wherein the level of T cells in the mesenteric lymph node is the level of Th memory cells in the mesenteric lymph node.

220. The method of any one of claims 216-218, wherein the level of T cells in the Peyer's patch is the level of Th memory cells in the Peyer's patch.

221. The method of any one of claims 216-218, wherein the level of T cells in the blood is the level of Th memory cells in the blood.

222. The method of any one of claims 216-218, wherein the topical administration of the immune modulator has been determined to result in a decrease in one or both of the level of T cells in a mesenteric lymph node and the level of T cells in a Peyer's patch in a mammal, each as compared to the corresponding level in a control mammal systemically administered the same dose of the immune modulator.

223. The method of any one of claims 216-218, wherein the topical administration of the immune modulator has been determined to result in an increase in the level of T cells in blood in the mammal, as compared to the corresponding level in a control mammal systemically administered the same dose of the immune modulator.

224. The method of any one of claims 216-218, wherein the topical administration of the immune modulator has been determined to result in (i) a decrease in one or both of the level of T cells in a mesenteric lymph node and the level of T cells in a Peyer's patch in a mammal, and (ii) an increase in the level of T cells in blood in the mammal, each as compared to the corresponding level in a control mammal systemically administered the same dose of the immune modulator.

225. The method of any one of claims 216-224, wherein the control mammal is a mammal of a similar age and having a similar disease state as compared to the mammal topically administered the dose of the immune modulator.

226. The method of any one of claims 216-225, wherein the immune modulator is selected from the group consisting of: IL-12/IL-23 inhibitors, TNF.alpha.
inhibitors, IL-6 receptor inhibitors, CD40/CD40L inhibitors, IL-1 inhibitors, IL-13 inhibitors, IL-10 receptor agonists, and integrin inhibitors.

227. The method of any one of claims 216-226, wherein the pharmaceutical composition is an ingestible device that contains a therapeutically effective amount of the immune modulator disposed therein.

228. A pharmaceutical composition prepared by the method of any one of claims 216-227.

229. A kit comprising the pharmaceutical composition of claim 228.